Benzodiazepine derivatives, preparation thereof and use thereof

ABSTRACT

Compounds of Formula (I) wherein R 1 , R 3 , R 4 , R 5  and X are as defined in the specification, as well as salts, enantiomers thereof and pharmaceutical compositions including the compounds are prepared. They are useful in therapy, in particular in the management of pain

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a US National Stage of International Application No. PCT/SE02/02306 that was filed on Dec. 11, 2002. The International Application claims priority under 35 U.S.C. § 119(a) to Swedish Application No. 0104250-6 filed Dec. 14, 2001.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention is directed to compounds that are useful in treating or preventing pain, septic shock, pancreatitis, edema, rhinitis, asthma, colitis, arthritis, hepatorenal syndrome, cancer, bacterial and viral infections, ulcerative colitis, and Alzheimer's Disease. More particularly, the present invention is directed to benzodiazepine derivatives that useful in treating pain.

2. Discussion of Relevant Art

Two types of bradykinin receptor are known: The B1 receptor and the B2 receptor. A number of reports indicate an important role for the B2 receptor in the pathophysiology of pain. [e.g. Hall, J. M., Morton, I. K. M. The pharmacology and immunopharmacology of kinin receptors. In: Farmer S G (Ed). The kinin system. London: Academic Press, 1997; 9-44]. Hence, compounds that are B2 antagonists are useful in the relief of pain, including chronic pain and acute pain, e.g., chronic inflammatory pain, neuropathic pain, back pain, migraine, cancer pain, visceral pain, arthritis pain and post-operative pain.

DETAILED DESCRIPTION OF THE INVENTION

Thus, the problem underlying the present invention was to develop new compounds that are novel kinin B2 antagonists.

Accordingly, in one aspect, the present invention provides compounds that are useful in treating pain.

Definitions

Unless specified otherwise within this specification, the nomenclature used in this specification generally follows the examples and rules stated in Nomenclature of Organic Chemistry, Sections A, B, C, D, E, F, and H, Pergamon Press, Oxford, 1979, which is incorporated by references herein for its exemplary chemical structure names and rules on naming chemical structures. Optionally, a name of a compound may be generated using a chemical naming program: ACD/ChemSketch, Version 5.09/September 2001, Advanced Chemistry Development, Inc., Toronto, Canada.

The term “C_(m-n)” or “C_(m-n) group” used alone or as a prefix, refers to any group having m to n carbon atoms, and having 0 to n multivalent heteroatoms selected from O, S, N and P, wherein m and n are O or positive integers, and n>m. For example, “C₁₋₆” would refer to a chemical group having 1 to 6 carbon atoms, and having 0 to 6 multivalent heteroatoms selected from O, S, N and P.

The term “hydrocarbon” used alone or as a suffix or prefix, refers to any structure comprising only carbon and hydrogen atoms up to 14 carbon atoms.

The term “hydrocarbon radical” or “hydrocarbyl” used alone or as a suffix or prefix, refers to any structure as a result of removing one or more hydrogens from a hydrocarbon.

The term “alkyl” used alone or as a suffix or prefix, refers to monovalent straight or branched chain hydrocarbon radicals comprising 1 to about 12 carbon atoms. Unless otherwise specified, “alkyl” general includes both saturated alkyl and unsaturated alkyl.

The term “alkylene” used alone or as suffix or prefix, refers to divalent straight or branched chain hydrocarbon radicals comprising 1 to about 12 carbon atoms, which serves to links two structures together.

The term “alkenyl” used alone or as suffix or prefix, refers to a monovalent straight or branched chain hydrocarbon radical having at least one carbon-carbon double bond and comprising at least 2 up to about 12 carbon atoms.

The term “alkynyl” used alone or as suffix or prefix, refers to a monovalent straight or branched chain hydrocarbon radical having at least one carbon-carbon triple bond and comprising at least 2 up to about 12 carbon atoms.

The term “cycloalkyl,” used alone or as suffix or prefix, refers to a monovalent ring-containing hydrocarbon radical comprising at least 3 up to about 12 carbon atoms.

The term “cycloalkenyl” used alone or as suffix or prefix, refers to a monovalent ring-containing hydrocarbon radical having at least one carbon-carbon double bond and comprising at least 3 up to about 12 carbon atoms.

The term “cycloalkenyl” used alone or as suffix or prefix, refers to a monovalent ring-containing hydrocarbon radical having at least one carbon-carbon triple bond and comprising about 7 up to about 12 carbon atoms.

The term “aryl” used alone or as suffix or prefix, refers to a monovalent hydrocarbon radical having one or more polyunsaturated carbon rings having aromatic character, (e.g., 4n+2 delocalized electrons) and comprising 5 up to about 14 carbon atoms.

The term “arylene” used alone or as suffix or prefix, refers to a divalent hydrocarbon radical having one or more polyunsaturated carbon rings having aromatic character, (e.g., 4n+2 delocalized electrons) and comprising 5 up to about 14 carbon atoms, which serves to links two structures together.

The term “heterocycle” used alone or as a suffix or prefix, refers to a ring-containing structure or molecule having one or more multivalent heteroatoms, independently selected from N, O, P and S, as a part of the ring structure and including at least 3 and up to about 20 atoms in the ring(s). Heterocycle may be saturated or unsaturated, containing one or more double bonds, and heterocycle may contain more than one ring. When a heterocycle contains more than one ring, the rings may be fused or unfused. Fused rings generally refer to at least two rings share two atoms therebetween. Heterocycle may have aromatic character or may not have aromatic character.

The term “heteroalkyl” used alone or as a suffix or prefix, refers to a radical formed as a result of replacing one or more carbon atom of an alkyl with one or more heteroatoms selected from N, O, P and S.

The term “heteroaromatic” used alone or as a suffix or prefix, refers to a ring-containing structure or molecule having one or more multivalent heteroatoms, independently selected from N, O, P and S, as a part of the ring structure and including at least 3 and up to about 20 atoms in the ring(s), wherein the ring-containing structure or molecule has an aromatic character (e.g., 4n+2 delocalized electrons).

The term “heterocyclic group,” “heterocyclic moiety,” “heterocyclic,” or “heterocyclo” used alone or as a suffix or prefix, refers to a radical derived from a heterocycle by removing one or more hydrogens therefrom.

The term “heterocyclyl” used alone or as a suffix or prefix, refers a monovalent radical derived from a heterocycle by removing one hydrogen therefrom.

The term “heterocyclylene” used alone or as a suffix or prefix, refers to a divalent radical derived from a heterocycle by removing two hydrogens therefrom, which serves to links two structures together.

The term “heteroaryl” used alone or as a suffix or prefix, refers to a heterocyclyl having aromatic character.

The term “heterocylcoalkyl” used alone or as a suffix or prefix, refers to a heterocyclyl that does not have aromatic character.

The term “heteroarylene” used alone or as a suffix or prefix, refers to a heterocyclylene having aromatic character.

The term “heterocycloalkylene” used alone or as a suffix or prefix, refers to a heterocyclylene that does not have aromatic character.

The term “six-membered” used as prefix refers to a group having a ring that contains six ring atoms.

The term “five-membered” used as prefix refers to a group having a ring that contains five ring atoms.

A five-membered ring heteroaryl is a heteroaryl with a ring having five ring atoms wherein 1, 2 or 3 ring atoms are independently selected from N, O and S.

Exemplary five-membered ring heteroaryls are thienyl, furyl, pyrrolyl, imidazolyl, thiazolyl, oxazolyl, pyrazolyl, isothiazolyl, isoxazolyl, 1,2,3-triazolyl, tetrazolyl, 1,2,3-thiadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-triazolyl, 1,2,4-thiadiazolyl, 1,2,4-oxadiazolyl, 1,3,4-triazolyl, 1,3,4-thiadiazolyl, and 1,3,4-oxadiazolyl.

A six-membered ring heteroaryl is a heteroaryl with a ring having six ring atoms wherein 1, 2 or 3 ring atoms are independently selected from N, O and S.

Exemplary six-membered ring heteroaryls are pyridyl, pyrazinyl, pyrimidinyl, triazinyl and pyridazinyl.

The term “substituted” used as a prefix refers to a structure, molecule or group, wherein one or more hydrogens are replaced with one or more C₁₋₁₂hydrocarbon groups, or one or more chemical groups containing one or more heteroatoms selected from N, O, S, F, Cl, Br, I, and P. Exemplary chemical groups containing one or more heteroatoms include heterocyclyl, —NO₂, —OR, —Cl, —Br, —I, —F, —CF₃, —C(═O)R, —C(═O)OH, —NH₂, —SH, —NHR, —NR₂, —SR, —SO₃H, —SO₂R, —S(═O)R, —CN, —OH, —C(═O)OR, —C(═O)NR₂, NRC(═O)R, oxo (═O), imino (═NR), thio (═S), and oximino (═N—OR), wherein each “R” is a C₁₋₁₂hydrocarbyl. For example, substituted phenyl may refer to nitrophenyl, pyridylphenyl, methoxyphenyl, chlorophenyl, aminophenyl, etc., wherein the nitro, pyridyl, methoxy, chloro, and amino groups may replace any suitable hydrogen on the phenyl ring.

The term “substituted” used as a suffix of a first structure, molecule or group, followed by one or more names of chemical groups refers to a second structure, molecule or group, which is a result of replacing one or more hydrogens of the first structure, molecule or group with the one or more named chemical groups. For example, a “phenyl substituted by nitro” refers to nitrophenyl.

The term “optionally substituted” refers to both groups, structures, or molecules that are substituted and those that are not substituted.

Heterocycle includes, for example, monocyclic heterocycles such as: aziridine, oxirane, thiirane, azetidine, oxetane, thietane, pyrrolidine, pyrroline, imidazolidine, pyrazolidine, pyrazoline, dioxolane, sulfolane 2,3-dihydrofuran, 2,5-dihydrofuran tetrahydrofuran, thiophane, piperidine, 1,2,3,6-tetrahydro-pyridine, piperazine, morpholine, thiomorpholine, pyran, thiopyran, 2,3-dihydropyran, tetrahydropyran, 1,4-dihydropyridine, 1,4-dioxane, 1,3-dioxane, dioxane, homopiperidine, 2,3,4,7-tetrahydro-1H-azepine homopiperazine, 1,3-dioxepane, 4,7-dihydro-1,3-dioxepin, and hexamethylene oxide.

In addition, heterocycle includes aromatic heterocycles, for example, pyridine, pyrazine, pyrimidine, pyridazine, thiophene, furan, furazan, pyrrole, imidazole, thiazole, oxazole, pyrazole, isothiazole, isoxazole, 1,2,3-triazole, tetrazole, 1,2,3-thiadiazole, 1,2,3-oxadiazole, 1,2,4-triazole, 1,2,4-thiadiazole, 1,2,4-oxadiazole, 1,3,4-triazole, 1,3,4-thiadiazole, and 1,3,4-oxadiazole.

Additionally, heterocycle encompass polycyclic heterocycles, for example, indole, indoline, isoindoline, quinoline, tetrahydroquinoline, isoquinoline, tetrahydroisoquinoline, 1,4-benzodioxan, coumarin, dihydrocoumarin, benzofuran, 2,3-dihydrobenzofuran, isobenzofuran, chromene, chroman, isochroman, xanthene, phenoxathiin, thianthrene, indolizine, isoindole, indazole, purine, phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline, pteridine, phenanthridine, perimidine, phenanthroline, phenazine, phenothiazine, phenoxazine, 1,2-benzisoxazole, benzothiophene, benzoxazole, benzthiazole, benzimidazole, benztriazole, thioxanthine, carbazole, carboline, acridine, pyrolizidine, and quinolizidine.

In addition to the polycyclic heterocycles described above, heterocycle includes polycyclic heterocycles wherein the ring fusion between two or more rings includes more than one bond common to both rings and more than two atoms common to both rings. Examples of such bridged heterocycles include quinuclidine, diazabicyclo[2.2.1]heptane and 7-oxabicyclo[2.2.1]heptane.

Heterocyclyl includes, for example, monocyclic heterocyclyls, such as: aziridinyl, oxiranyl, thiiranyl, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrrolinyl, imidazolidinyl, pyrazolidinyl, pyrazolinyl, dioxolanyl, sulfolanyl, 2,3-dihydrofuranyl, 2,5-dihydrofuranyl, tetrahydrofuranyl, thiophanyl, piperidinyl, 1,2,3,6-tetrahydro-pyridinyl, piperazinyl, morpholinyl, thiomorpholinyl, pyranyl, thiopyranyl, 2,3-dihydropyranyl, tetrahydropyranyl, 1,4-dihydropyridinyl, 1,4-dioxanyl, 1,3-dioxanyl, dioxanyl, homopiperidinyl, 2,3,4,7-tetrahydro-1H-azepinyl, homopiperazinyl, 1,3-dioxepanyl, 4,7-dihydro-1,3-dioxepinyl, and hexamethylene oxidyl.

In addition, heterocyclyl includes aromatic heterocyclyls or heteroaryl, for example, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, thienyl, furyl, furazanyl, pyrrolyl, imidazolyl, thiazolyl, oxazolyl, pyrazolyl, isothiazolyl, isoxazolyl, 1,2,3-triazolyl, tetrazolyl, 1,2,3-thiadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-triazolyl, 1,2,4-thiadiazolyl, 1,2,4-oxadiazolyl, 1,3,4-triazolyl, 1,3,4-thiadiazolyl, and 1,3,4 oxadiazolyl.

Additionally, heterocyclyl encompasses polycyclic heterocyclyls (including both aromatic or non-aromatic), for example, indolyl, indolinyl, isoindolinyl, quinolinyl, tetrahydroquinolinyl, isoquinolinyl, tetrahydroisoquinolinyl, 1,4-benzodioxanyl, coumarinyl, dihydrocoumarinyl, benzofuranyl, 2,3-dihydrobenzofuranyl, isobenzofuranyl, chromenyl, chromanyl, isochromanyl, xanthenyl, phenoxathiinyl, thianthrenyl, indolizinyl, isoindolyl, indazolyl, purinyl, phthalazinyl, naphthyridinyl, quinoxalinyl, quinazolinyl, cinnolinyl, pteridinyl, phenanthridinyl, perimidinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxazinyl, 1,2-benzisoxazolyl, benzothiophenyl, benzoxazolyl, benzthiazolyl, benzimidazolyl, benztriazolyl, thioxanthinyl, carbazolyl, carbolinyl, acridinyl, pyrolizidinyl, and quinolizidinyl.

In addition to the polycyclic heterocyclyls described above, heterocyclyl includes polycyclic heterocyclyls wherein the ring fusion between two or more rings includes more than one bond common to both rings and more than two atoms common to both rings. Examples of such bridged heterocycles include quinuclidinyl, diazabicyclo[2.2.1]heptyl; and 7-oxabicyclo[2.2.1]heptyl.

The term “alkoxy” used alone or as a suffix or prefix, refers to radicals of the general formula —O—R, wherein R is selected from a hydrocarbon radical. Exemplary alkoxy includes methoxy, ethoxy, propoxy, isopropoxy, butoxy, t-butoxy, isobutoxy, cyclopropylmethoxy, allyloxy, and propargyloxy.

The term “amine” or “amino” used alone or as a suffix or prefix, refers to radicals of the general formula —NRR′, wherein R and R′ are independently selected from hydrogen or a hydrocarbon radical.

“Acyl” used alone, as a prefix or suffix, means —C(═O)—R, wherein R is an optionally substituted hydrocarbyl, hydrogen, amino or alkoxy. Acyl groups include, for example, acetyl, propionyl, benzoyl, phenyl acetyl, carboethoxy, and dimethylcarbamoyl.

Halogen includes fluorine, chlorine, bromine and iodine.

“Halogenated,” used as a prefix of a group, means one or more hydrogens on the group is replaced with one or more halogens.

“RT” or “rt” means room temperature.

A first ring group being “fused” with a second ring group means the first ring and the second ring share at least two atoms therebetween.

“Link,” “linked,” or “linking,” unless otherwise specified, means covalently linked or bonded.

Description of Preferred Embodiments

In one aspect, the present invention provides a compound of formula (I), pharmaceutically acceptable salts thereof, diasteriomers thereof, enantiomers thereof, or mixtures thereof:

wherein

R¹ is selected from optionally substituted acyl, optionally substituted alkyl-oxycarbonyl, optionally substituted alkyl, optionally substituted heteroalkyl, optionally substituted cycloalkyl, optionally substituted aryl; optionally substituted heterocyclyl; optionally substituted aryl-C₁₋₆alkyl, and optionally substituted heterocyclyl-C₁₋₆alkyl; or a divalent C₁₋₁₂ group that together with a second nitrogen of X to form a ring;

X is a divalent group including a first nitrogen atom and the second nitrogen atom, wherein a first group (e.g., the 2H,1,4benzodiazepin-2-one group of formula (I)) is linked to the first nitrogen atom and R¹ is linked to the second nitrogen atom, and wherein the first and second nitrogen atoms are separated by either one carbon atom, or two carbon atoms wherein said two carbon atoms have a double bond therebetween;

R³ is optionally substituted aryl, optionally substituted C₁₋₁₂alkyl, optionally substituted C₃₋₁₂cycloalkyl, or optionally substituted heterocyclyl;

R⁴ is, at each position, independently —H, halogen, optionally substituted alkyl, optionally substituted heteroalkyl, nitro, cyano, hydroxy, —OR⁶, —SR⁶, —S(═O)R⁶, —S(═O)₂R⁶, —C(═O)R⁶, —C(═S)R⁶, —NR⁷R⁶, —C(═O)NR⁷R⁶, —NR⁷C(═O)R⁶, —SO₂NR⁷R⁶, —NR⁷SO₂R⁶, or —C(═O)OR⁶; and

R⁵, R⁶ and R⁷ are independently —H, optionally substituted C₁₋₆alkyl.

In another aspect, the compounds of the present invention are those of formula (I), pharmaceutically acceptable salts thereof, diasteriomers thereof, enantiomers thereof, or mixtures thereof, wherein

R¹ is selected from optionally substituted acyl, optionally substituted alkyl-oxycarbonyl, optionally substituted alkyl, optionally substituted heteroalkyl, optionally substituted cycloalkyl, optionally substituted aryl; optionally substituted heterocyclyl; optionally substituted aryl-C₁₋₆alkyl, and optionally substituted heterocyclyl-C₁₋₆alkyl; or a divalent C₁₋₁₂ group that together with a divalent R² of X forms a portion of a ring;

X is represented by (i), (ii), (iii), (iv), (v), (vi), (vii), (viii), (ix), (x), (xi), (xii), (xiii), (xiv), (xv), (xvi), or (xvii) below:

wherein R² is selected from —H, optionally substituted C₁₋₁₂alkyl, optionally substituted C₁₋₁₂heteroalkyl, optionally substituted aryl, optionally substituted heterocyclyl, and a divalent C₀₋₆group together with a divalent R¹ to form the portion of the ring, wherein said divalent C₀₋₆ group optionally includes one or more heteroatoms;

R³ is optionally substituted aryl, optionally substituted C₁₋₁₂alkyl, optionally substituted C₃₋₁₂cycloalkyl, or optionally substituted heterocyclyl;

R⁴ is, at each position, independently, —H, halogen, optionally substituted alkyl, optionally substituted heteroalkyl, nitro, cyano, hydroxy, —OR⁶, —SR⁶, —S(═O)R⁶, —S(═O)₂R⁶, —C(═O)R⁶, —C(═S)R⁶, —NR⁷R⁶, —C(═O)NR⁷R⁶, —NR⁷C(═O)R⁶, —SO₂NR⁷R⁶, —NR⁷SO₂R⁶, or —C(═O)OR⁶; and

R⁵, R⁶ and R⁷ are independently —H, optionally substituted C₁₋₆alkyl.

More particularly, the compound of the present invention is a compound of formula (I), wherein

R¹ is optionally substituted phenyl, optionally substituted naphthyl, optionally substituted isoquinolyl, optionally substituted acridinyl, optionally substituted coumarinyl, optionally substituted carbazolyl, or a first divalent group selected from optionally substituted C₁₋₁₂alkylene and optionally substituted C₁₋₁₂heteroalkylene; wherein said phenyl, naphthyl, isoquinolyl, acridinyl, coumarinyl, and carbazolyl are optionally substituted by C₁₋₆alkyl, C₁₋₆heterocyclyl or amino, wherein said C₁₋₁₂alkylene and C₁₋₁₂heteroalkylene are optionally substituted by C₁₋₆alkyl, aryl-C₁₋₆alkyl, aryl or heterocyclyl;

X is selected from formulas (i), (ii), (iii), (vi) and (xvii) below:

R² is —H, C₁₋₃alkyl, or a second divalent group selected from a single bond, an optionally substituted alkylene and an optionally substituted heteroalkylene; wherein said second divalent group together with said first divalent group forms a portion of a ring;

R³ is optionally substituted aryl, optionally substituted heteroaryl or optionally substituted cycloalkyl;

R⁴ is halogen, or C₁₋₃alkyl; and

R⁵ is C₁₋₃alkyl.

Most particularly, the compound of the present invention is a compound of formula (I), wherein

—X—R¹ of formula (I) is selected from formulas (a), (b), (c), (d), (e), (f) and (g) below:

R¹ is optionally substituted phenyl, optionally substituted naphthyl, optionally substituted isoquinolyl, wherein said phenyl, naphthyl and isoquinolyl are optionally substituted by C₁₋₆alkyl, C₁₋₆heterocyclyl or amino;

R² is —H, or C₁₋₃alkyl;

is a nitrogen containing heterocyclyl, which may be optionally substituted by one or more —R⁸, and which includes a bond on the nitrogen that links to other group of formula (I). Exemplary nitrogen containing heterocyclyls include, but is not limited to, piperazinyl, morpholinyl, poperidyl, and pyrrolidinyl.

R⁸ is —H, optionally substituted aryl, optionally substituted heterocyclyl, optionally substituted C₁₋₆alkyl, —OH, or C₁₋₆alkoxy, wherein R⁸ is optionally fused with the ring of

R³ is optionally substituted cyclohexyl, optionally substituted phenyl, optionally substituted pyridyl, optionally substituted thienyl, or optionally substituted pyrimidinyl, wherein said cyclohexyl, phenyl, pyridyl, thienyl and pyrimidinyl are optionally substituted by halogen, methoxy, or C₁₋₃alkyl;

R⁴ is halogen; and

R⁵is methyl.

Specific examples of compounds of the present invention that may be used in practicing the present invention are listed in Table 1, below.

TABLE 1 Spreadsheet of combinatorially prepared compounds with LCMS analysis of the reaction product. Compound Target Mass Rtn Time # Structure Mass Found? (MS) 1

474.13 Yes 3.92 2

471.15 Yes 2.9 3

449.11 Yes 2.93 4

466.13 No 5

483.15 Yes 2.85 6

517.17 Yes 3.97 7

627.16 No 8

525.2 Yes 3.47 9

477.14 Yes 3.11 10

503.15 No 11

543.19 Yes 4.24 12

468.08 Yes 3.55 13

531.19 No 14

455.15 Yes 3.11 15

463.12 Yes 3.05 16

449.11 Yes 2.94 17

478.12 Tentative 18

449.11 Yes 2.89 19

552.17 Yes 4.5 20

469.17 Yes 2.85 21

469.13 Yes 2.78 22

499.14 Yes 3.3 23

586.14 Yes 3.75 24

532.17 Yes 3.7 25

561.16 Yes 3.87 26

477.14 Yes 3.17 27

469.17 Yes 2.78 28

509.2 Yes 2.74 29

504.15 Yes 3.43 30

502.13 Yes 2.96 31

571.14 Yes 32

509.2 Yes 4.05 33

629.18 Yes 34

571.08 No 35

531.19 Yes 4.8 36

565.19 No 37

533.17 Yes 3.87 38

547.18 Yes 3.98 39

521.15 Yes 4.02 40

513.16 Yes 3.22 41

593.2 Yes 42

455.15 Yes 2.79 43

428.11 Yes 2.91 44

495.19 Yes 2.62 45

538.16 Yes 4.64 46

505.15 Yes 3.31 47

457.17 Yes 3.02 48

477.14 Yes 3.17 49

571.14 Yes 4.49 50

471.19 Yes 2.9 51

521.15 Yes 4.02 52

519.19 Yes 3.97 53

545.17 Yes 3.49 54

441.14 Yes 2.94 55

521.15 Yes 3.92 56

477.14 Yes 3.08 57

456.14 Yes 3.34 58

492.14 Yes 3.47 59

521.13 Yes 3.04 60

497.2 Yes 2.7 61

452.12 Yes 2.69 62

443.15 Yes 2.51 63

513.14 Yes 3.97 64

483.19 Tentative 1.63 65

455.15 Yes 2.8 66

455.15 Yes 2.77 67

469.13 Yes 2.69 68

515.15 Yes 3.59 69

515.15 Yes 3.59 70

469.13 Yes 2.73 71

456.14 Tentative 72

471.15 Tentative 73

442.12 No 74

519.15 Tentative 75

533.17 No 76

538.19 No 77

512.21 No 78

525.16 Yes 6.5 79

574.19 No 80

504.15 Tentative 81

505.15 No 82

495.19 No 83

571.08 No 84

531.19 Yes 4.18 85

469.17 Yes 3.28 86

616.13 No 87

455.15 Yes 88

503.15 Yes 5.38 89

483.15 No 90

622.15 Yes 6.44 91

571.14 No 92

620.1 Yes 4.37 93

531.19 No 94

531.19 No 95

428.11 Tentative 96

572.14 Tentative 6.45 97

518.13 No 98

661.12 Yes 99

613.23 Yes 6.59 100

532.17 No 101

558.16 No 102

598.19 No 103

519.15 No 104

470.15 Yes 105

456.14 Yes 106

463.12 Yes 107

463.12 Yes 5.94 108

469.17 Yes 2.7 109

485.17 No 110

498.2 No 111

513.16 Tentative 112

512.14 Yes 4.21 113

469.17 Yes 2.55 114

512.14 No 115

469.17 No 116

504.17 No 117

476.08 Tentative 118

490.12 No 119

457.17 Yes 2.68 120

499.22 No 121

455.15 Yes 2.63 122

497.2 No 123

493.09 Yes 3.34 124

469.17 No 125

467.15 No 126

443.15 No 127

469.17 No 128

505.17 No 129

569.22 No 130

555.21 Yes 3.99 131

555.21 Yes 4.03 132

529.19 Yes 3.74 133

527.18 No 134

541.19 No 135

541.19 Yes 3.84 136

527.18 Tentative 137

541.19 Yes 3.84 138

599.2 Yes 3.87 139

541.19 Yes 3.75 140

539.18 No 141

599.2 Yes 3.87 142

572.18 No 143

556.14 No 144

558.16 No 145

501.16 No 146

577.19 No 147

527.18 Yes 3.46 148

587.2 No 149

541.19 Yes 3.59 150

474.13 No 151

466.1 Yes 3.54 152

541.1 Yes 2.63, 2.81 153

553.17 Tentative 4.13 154

412.11 Tentative 2.21 155

471.15 Yes 2.9 156

453.14 Yes 3.37, 4.15 157

441.14 Yes 2.61 158

442.12 No 159

470.19 Yes 4.55, 4.66 160

426.13 Yes 3.27 161

504.17 Yes 4.52 162

536.16 Yes 3.46 163

476.14 Yes 3.97 164

562.16 Yes 3.1, 3.35 165

462.13 Yes 3.64 166

414.13 No 167

460.13 Yes 3.08 168

427.12 Yes 2.63 169

416.11 Yes 2.7 170

440.14 Yes 3.59 171

454.16 No 172

398.1 No 173

538.16 Yes 4.35 174

474.15 Yes 3.23 175

424.11 Yes 3.66 176

484.13 Yes 3.13 177

454.16 Yes 3.87 178

498.15 Yes 3.44 179

438.09 Yes 2.66, 3.16 180

480.12 Yes 3.68 181

492.14 Yes 3.44 182

494.19 Yes 3.68, 4.04,4.78 183

440.14 Yes 3.5 184

440.14 Yes 3.59 185

414.13 No 186

454.16 No 187

410.1 Tentative 2.81, 3.33 188

410.1 Yes 3.03, 6.5 189

492.14 Yes 3.59 190

494.19 Yes 4.73 191

548.14 Yes 3.62 192

498.13 Yes 4.06 193

456.14 Yes 3.21 194

492.1 Yes 3.4 195

476.14 Yes 3.88 196

501.14 Yes 3.22, 3.43 197

512.14 Yes 4.26 198

474.13 Yes 3.83 199

412.11 Tentative 3.12 200

498.15 Yes 3.39 201

458.12 Yes 2.96 202

532.09 Yes 4.16 203

538.14 Yes 3.24 204

440.07 No 205

516.17 Yes 4.45 206

608.2 Yes 4.22 207

443.15 Yes 2.57 208

517.17 Yes 3.73 209

474.15 Yes 3.39 210

466.16 Yes 4.22 211

466.16 Yes 3.99 212

508.13 Yes 3.19 213

524.14 Yes 4.17 214

538.16 Yes 4.39 215

492.14 Yes 3.64 216

463.12 Yes 2.56, 2.82 217

552.17 Yes 4.82 218

516.12 Yes 3.37 219

534.09 Yes 4.24 220

428.14 Yes 3.61 221

478.12 Yes 3.41 222

540.04 Yes 4.21 223

500.13 Yes 3.08 224

472.13 Tentative 3.13 225

502.16 No 226

506.19 Yes 4.8 227

438.13 Yes 3.52 228

490.16 Yes 4.19 229

440.14 Yes 3.52 230

522.15 Yes 3.32 231

478.12 Yes 3.49 232

396.08 No 233

458.12 Yes 3.12 234

474.13 No 235

482.07 Yes 3.29, 3.86 236

464.14 Yes 2.68, 4.77 237

442.12 Yes 3.02 238

398.1 No 239

502.18 Tentative 4.31 240

508.12 Yes 3.99 241

496.09 Yes 4.09 242

446.1 Yes 3.38 243

471.19 Yes 2.86 244

529.15 Yes 3.41 245

572.14 Yes 4.16 246

538.11 Yes 4 247

526.02 No 248

468.17 No 249

518.17 Yes 3.85 250

492.14 Yes 3.74 251

516.1 Yes 3.9 252

462.13 Yes 3.68 253

430.12 Yes 3.03 254

443.15 Yes 2.77 255

426.13 Yes 3.4 256

400.11 No 257

466.1 Yes 2.83, 3.54 258

416.11 Yes 2.86 259

448.11 Yes 3.45 260

600.06 Yes 2.43, 3.84 261

626.28 Yes 6.29 262

386.1 No 263

425.11 Yes 2.84 264

506.15 Yes 4.05 265

456.14 Yes 2.77 266

454.16 Yes 3.79 267

454.16 Yes 3.97 268

444.14 Yes 3.28 269

444.1 Yes 2.8 270

476.14 Yes 3.86 271

456.14 Yes 3.32 272

442.16 Yes 3.8 273

470.19 Yes 4.39 274

441.14 Yes 2.93 275

454.07 Yes 3.35 276

466.13 Yes 2.81 277

442.12 Yes 3.01 278

456.14 Tentative 3.13 279

508.13 Yes 3.48 280

430.12 Yes 2.95 281

482.07 Yes 3.89 282

428.14 Yes 3.53 283

412.11 Yes 3.19 284

454.16 Yes 3.84 285

478.12 Yes 3.03, 3.16,4.05 286

538.16 Yes 4.6 287

476.14 Yes 3.87 288

440.14 Yes 3.68 289

442.16 Yes 3.91 290

456.14 Yes 3.16 291

455.06 No 292

414.13 Tentative 3.12 293

518.15 Yes 3.51 294

466.1 Yes 3.5 295

470.15 No 296

494.13 Yes 3.99 297

440.14 Yes 3.61 298

398.1 No 299

452.11 No 300

452.11 No 301

442.12 Yes 3.02 302

452.11 Yes 2.78, 3.41 303

416.11 Yes 2.68 304

458.06 No 305

512.14 Yes 4.17 306

478.12 Yes 3.01 307

458.12 Yes 2.86 308

454.16 Yes 4.01 309

456.14 Yes 2.77 310

476.14 Yes 3.8 311

412.11 Yes 3.18, 6.2 312

476.07 Yes 2.75 313

492.06 Yes 3.77 314

484.09 Yes 3.45 315

476.14 Yes 3.9 316

492.14 Yes 3.77 317

454.16 Yes 3.91 318

517.17 No 319

586.14 Yes 4 320

490.12 Yes 3.41, 3.63 321

439.09 Yes 3.33 322

446.12 Yes 2.96 323

472.13 Yes 3.14 324

476.14 Yes 3.99 325

494.19 No 326

506.12 Yes 3.83 327

443.12 Yes 2.68 328

464.12 Yes 2.92 329

535.18 Yes 2.99 330

516.1 Yes 4.03 331

529.19 Yes 3.51 332

555.21 Yes 3.91, 4.15 333

517.1 Yes 3.44 334

531.19 Yes 3.6, 3.76 335

573.16 Yes 4.27 336

615.17 Yes 4.19 337

539.13 Yes 3.94 338

565.15 Yes 4.19, 4.36 339

581.14 Yes 3.9 340

535.18 Yes 3.43 341

488.12 No 342

527.09 Yes 2.73 343

469.17 Yes 2.58 344

530.05 Yes 4.59 345

480.12 Yes 3.73 346

480.12 Yes 3.06, 3.76 347

573.16 Yes 3.02 348

518.19 Yes 4.85 349

531.15 Yes 2.69, 3,3.25, 3.36 350

477.14 Yes 2.94 351

496.09 Yes 4.02 352

531.15 Yes 2.59, 3.2,3.31 353

559.22 Yes 3.37 354

456.14 Yes 3.01 355

442.12 Yes 2.97 356

470.15 Yes 3.15 357

496.09 Yes 4.11 358

471.19 Yes 2.6 359

515.15 Yes 3.49, 3.69 360

476.14 Yes 4.01 361

425.08 Yes 2.7 362

452.12 Yes 2.94 363

497.11 Yes 4.37 364

438.1 Yes 3.45 365

441.05 Yes 3.27 366

455.06 Yes 3.49 367

471.07 No 368

509.05 No 369

424.09 Yes 3.15 370

508.09 No 371

500.06 Yes 3.59 372

575.06 Yes 2.87 373

587.13 No 374

446.07 No 375

505.11 Yes 2.96 376

487.1 Yes 3.29, 4.02 377

475.1 Yes 2.68 378

476.08 No 379

504.15 Yes 4.64 380

460.09 Yes 3.29 381

538.14 Yes 4.51 382

570.13 Yes 3.49 383

510.1 Yes 3.96 384

596.12 Yes 385

496.09 Yes 3.66 386

448.09 No 387

494.09 Yes 3.12 388

461.08 Yes 2.71 389

450.07 Yes 2.77 390

474.1 Yes 3.6 391

488.12 No 392

432.06 No 393

572.12 Yes 4.36 394

508.11 Yes 3.24 395

458.07 Yes 3.55 396

518.09 Yes 3.16 397

488.12 Yes 3.88 398

532.11 Yes 3.44 399

472.05 Yes 3.21 400

514.08 Yes 3.72 401

526.1 Yes 3.46 402

528.15 No 403

474.1 Yes 3.51 404

474.1 Yes 3.59 405

448.09 No 406

488.12 No 407

444.06 Yes 2.84, 3.26 408

444.06 Yes 3.06 409

526.1 Yes 3.64 410

528.15 Yes 4.73 411

582.1 Yes 3.63 412

532.09 Yes 4.08 413

490.1 Yes 3.23 414

526.06 Yes 3.44 415

510.1 Yes 3.92 416

535.1 Yes 2.86, 3.28,3.48 417

546.1 Yes 4.25 418

508.09 Yes 3.81 419

446.07 Tentative 3.1 420

532.11 Yes 3.41 421

492.08 Yes 3.03 422

566.06 Yes 4.19 423

572.11 Yes 3.3 424

474.03 No 425

550.14 Yes 4.43 426

642.16 Yes 4.22 427

477.12 Yes 2.65 428

551.13 Yes 3.74 429

508.11 Yes 3.42 430

500.12 Yes 4.24 431

500.12 Yes 4 432

542.09 Yes 3.25 433

558.1 Yes 4.2 434

572.12 Yes 4.4 435

526.1 Yes 3.69 436

497.08 Yes 2.68 437

586.14 Yes 4.83 438

550.08 Yes 3.41 439

568.05 Yes 4.27 440

462.1 Yes 3.66 441

512.08 Yes 3.46 442

574 Yes 4.24 443

534.09 Yes 3.14 444

506.09 Yes 3.24 445

536.12 No 446

540.15 Yes 4.79 447

472.09 Yes 3.55 448

524.12 Yes 4.19 449

474.1 Yes 3.53 450

556.11 Yes 3.38 451

512.08 Yes 3.52 452

430.04 Yes 3.11, 6.48 453

492.08 Yes 3.16 454

508.09 Yes 3.9 455

516.03 Yes 3.9 456

498.1 Yes 4.65, 6.23 457

476.08 Yes 3.08 458

432.06 No 459

536.14 No 460

542.08 Yes 3.25, 4.03 461

530.05 Yes 4.12 462

480.06 Yes 2.81, 3.43 463

505.15 Yes 2.93 464

563.11 Yes 3.43 465

606.1 Yes 4.14 466

572.07 Yes 3.96 467

559.98 Yes 3.86 468

502.14 No 469

552.13 Yes 3.83 470

526.1 Yes 3.75 471

550.06 Yes 3.92 472

496.09 Yes 3.72 473

464.08 Yes 3.08 474

477.12 Yes 2.85 475

460.09 Yes 3.44 476

434.07 No 477

500.06 Yes 3.59 478

450.07 Yes 2.93 479

482.07 Yes 3.49 480

634.02 Yes 3.89 481

660.25 No 482

420.06 No 483

459.07 Yes 2.89 484

540.12 Yes 4.07 485

490.1 Yes 2.85 486

488.12 Yes 1.84, 3.8 487

488.12 Yes 3.93 488

478.1 Yes 3.32 489

478.06 Yes 2.87 490

510.1 Yes 3.87 491

490.1 Yes 3.3 492

476.12 Yes 3.81 493

504.15 Yes 4.39 494

475.1 Yes 2.99 495

488.03 Yes 3.37 496

500.1 Yes 2.88 497

476.08 Yes 3.08 498

490.1 No 499

542.09 Yes 3.53 500

464.08 Yes 3 501

516.03 Yes 3.92 502

462.1 Yes 3.58 503

446.07 Yes 3.24 504

488.12 Yes 3.86 505

512.08 Yes 3.16 506

572.12 Yes 4.56 507

510.1 Yes 3.89 508

474.1 Yes 3.7 509

476.12 Yes 3.94 510

490.1 Yes 3.21 511

489.03 No 512

448.09 Tentative 2.76 513

552.12 Yes 3.53 514

500.06 Yes 3.54 515

504.12 Yes 3.34 516

528.1 Yes 3.98 517

474.1 Yes 3.61 518

432.06 No 519

486.07 No 520

486.07 No 521

476.08 Yes 3.07 522

486.07 Yes 3.46 523

450.07 Yes 2.75 524

492.02 Tentative 6.5 525

546.1 Yes 4.2 526

512.08 Yes 3.08 527

492.08 Yes 2.93 528

488.12 Yes 4.04 529

490.1 Yes 2.85 530

510.1 Yes 3.84 531

446.07 Yes 3.3 532

510.04 Yes 2.82 533

526.02 Yes 3.76 534

518.05 Yes 3.47 535

510.1 Yes 3.91 536

526.1 Yes 3.8 537

488.12 Yes 3.93 538

551.13 No 539

620.1 Yes 4.03 540

524.08 Yes 3.57 541

473.05 Yes 2.79, 3.36 542

480.08 Yes 3.02 543

506.09 Yes 3.19 544

510.1 Yes 4.02 545

528.15 No 546

540.08 Yes 3.86 547

477.08 Yes 2.76 548

498.08 Yes 2.97 549

569.14 Yes 3.06 550

550.06 Yes 4.06 551

563.15 Yes 3.57 552

589.17 Yes 3.89, 4.18 553

551.06 Yes 3.49 554

565.15 Yes 3.68 555

607.12 Yes 4.24 556

649.13 Yes 4.2 557

573.09 Yes 3.92 558

599.11 Yes 4.3 559

615.1 Yes 3.93 560

569.14 Yes 3.46 561

522.08 Yes 3.32 562

561.05 Yes 2.79 563

503.13 Tentative 2.68 564

564.01 Yes 4.61 565

514.08 Yes 3.75 566

514.08 Yes 3.8 567

607.12 Yes 3.86 568

552.15 Yes 4.85 569

565.11 Yes 3.41 570

511.1 Yes 3 571

530.05 Yes 4.03 572

565.11 Yes 3.36 573

593.18 Yes 3.43 574

490.1 Yes 3.08 575

476.08 Yes 3.03 576

504.12 Yes 3.21 577

530.05 Yes 4.14 578

505.15 Yes 2.69 579

549.12 Yes 3.54, 3.74 580

510.1 Yes 4.03 581

472.06 Yes 2.87 582

483.19 Yes 2.57 583

477.1 Yes 2.38, 2.62 584

469.2 Yes 2.63, 5.03 585

401.11 Yes 2.45 586

477.1 Yes 2.67, 3.16 587

473.13 No 588

455.2 Yes 2.56, 3.84 589

429.14 Yes 2.48 590

427.12 Yes 2.49 591

441.14 Yes 2.5 592

441.1 Yes 2.48, 2.92 593

427.12 Yes 2.62 594

441.14 Yes 2.58 595

499.1 Yes 2.81, 3.84 596

427.12 Yes 2.53 597

441.14 Yes 2.59 598

487.14 No 599

439.12 Yes 2.56 600

499.14 Yes 2.81 601

441.14 Yes 2.53 602

497.2 No 603

491.15 Tentative 2.87 604

483.19 Tentative 2.63 605

415.1 Yes 2.49, 3.01 606

491.15 No 607

487.14 No 608

469.17 Yes 2.64 609

443.15 Yes 610

441.14 Yes 2.76 611

455.15 Yes 2.68 612

455.15 Yes 2.76 613

441.14 Yes 2.93 614

455.15 Yes 2.8 615

513.16 Yes 3.13 616

441.14 Tentative 2.9 617

455.15 Yes 2.9 618

501.16 No 619

453.14 Yes 2.73 620

513.16 Yes 3.13 621

455.15 No 622

525.23 Yes 3.15 623

519.19 Yes 2.72 624

511.22 Yes 3.19 625

443.15 Yes 2.64 626

519.19 Tentative 627

515.18 Tentative 3.73 628

497.2 Yes 2.66, 2.88 629

471.19 Yes 2.76 630

469.17 Yes 2.76 631

483.19 Yes 2.68 632

483.19 Yes 2.78 633

469.2 Yes 2.51, 3.27 634

483.2 Yes 2.54, 2.91 635

541.19 Yes 3.79 636

469.2 Yes 2.76, 3.1 637

483.2 Yes 2.5, 3.09 638

529.19 No 639

481.2 Yes 2.5, 2.97 640

541.2 Yes 3.79, 4.07 641

483.2 Tentative 2.4, 2.98 642

524.21 Yes 2.82 643

518.17 Yes 2.64 644

510.2 Tentative 2.9 645

442.13 Yes 2.48 646

518.17 Yes 2.68 647

514.16 No 648

496.18 Yes 2.6 649

470.17 Yes 2.52 650

468.2 Yes 2.24, 2.53 651

482.2 Yes 2.15, 2.52 652

482.17 Yes 2.65 653

468.15 Yes 2.52 654

482.17 Yes 2.55 655

540.17 Yes 2.66 656

468.15 Yes 2.52 657

482.17 Yes 2.52 658

528.17 No 659

480.15 Yes 2.51 660

540.17 Yes 661

482.17 Yes 2.58 662

492.16 Yes 3.63 663

468.12 Yes 2.99 664

537.15 Yes 3.64 665

430.15 Yes 2.78 666

468.06 Tentative 3.33 667

468.06 Yes 3.76 668

460.16 Yes 2.76 669

468.06 Yes 670

430.15 No 671

428.17 Yes 3.08 672

445.12 Yes 3.03 673

478.05 No 674

425.13 Yes 2.89 675

458.14 No 676

471.21 Yes 3.41 677

444.16 Yes 3.01 678

460.16 Yes 2.68 679

434.1 Tentative 3.32 680

418.13 Yes 3.01 681

468.12 Yes 3.44 682

434.1 Yes 3.21, 6.45 683

434.1 Yes 2.93, 6.48 684

444.13 Yes 2.73 685

493.19 Yes 3.31 686

430.15 Yes 2.79 687

506.18 Yes 3.7 688

536.05 Yes 4.46 689

526.1 Yes 2.53, 4.15 690

502.08 Yes 3.39 691

571.11 No 692

464.11 Yes 3.16 693

502.02 Yes 3.81 694

502.02 Yes 695

494.1 Yes 3.14, 3.45 696

502.02 No 697

464.11 Yes 3.34 698

462.13 Yes 3.49 699

479.08 No 700

512.01 Yes 3.36 701

459.09 Yes 3.27 702

492.1 Yes 3.37, 6.53 703

505.17 Yes 3.86 704

478.12 Yes 3.46 705

494.12 Yes 2.99 706

468.06 Yes 3.76 707

452.09 Yes 3.45 708

502.08 Yes 3.92 709

468.06 Yes 3.74 710

468.06 Yes 3.34 711

478.1 Yes 3.12, 3.49 712

527.2 Yes 3.76, 4.81 713

464.1 Yes 3.19, 3.53 714

540.1(blank) Tentative 6.4 715

508.13 Yes 4.35, 6.57 716

498.21 Yes 4.18 717

474.17 Yes 3.4 718

543.19 Yes 2.52, 4.19 719

436.19 Yes 2.7, 3.18 720

474.1 Yes 3.76 721

474.1 Yes 4.21 722

466.2 Yes 3.2 723

474.1 Yes 3.96 724

436.19 Yes 3.4 725

434.21 Yes 3.57 726

451.17 Yes 3.45 727

484.09 Yes 3.37 728

431.18 Yes 3.23 729

464.19 Yes 3.34 730

477.26 Yes 3.98 731

450.21 Yes 3.48 732

466.2 Yes 3.04 733

440.14 Yes 3.72 734

424.17 Yes 3.41 735

474.17 Yes 3.87 736

440.14 Yes 3.7 737

440.14 Yes 3.35 738

450.17 Yes 3.09 739

499.24 Yes 3.87 740

436.19 Yes 3.18 741

512.22 No 742

449.11 No 743

498.07 Yes 3.74 744

581.11 No 745

482.07 Yes 4 746

436.09 Yes 2.98 747

492.1 Yes 3.1 748

518.07 Yes 749

485.11 Yes 3.02 750

485.11 Yes 3.63 751

499.12 Yes 3.42 752

488.14 Yes 3.86 753

469.05 Yes 3.4 754

516.1 No 755

459.09 Yes 3.27 756

536.11 Yes 3.51 757

490.16 Yes 4.28 758

612.14 Yes 2.89 759

448.11 Yes 3.44 760

546.22 Tentative 6.44 761

532.09 Yes 4.11 762

526.12 No 763

526.12 Yes 4.19 764

502.08 Yes 4.11 765

482.07 Yes 3.67 766

526.12 Yes 3.31 767

483.07 Yes 3.44 768

526.02 Yes 4.15 769

506.12 No 770

473.11 No 771

522.13 Yes 3.06 772

483.07 No 773

526.12 Yes 3.37 774

559.99 Yes 3.93 775

510.13 Yes 4.2 776

534.15 Yes 4.55 777

462.13 Yes 778

600.04 No 779

509.11 Yes 3.57 780

524.14 Yes 3.92 781

550.11 Yes 3.55 782

478.12 No 783

473.11 Tentative 2.93 784

549.14 Yes 3.87 785

550.11 Yes 3.57 786

514.12 Yes 787

434.1 Yes 3.21, 6.52 788

452.09 Yes 3.45 789

464.11 Yes 3.27, 3.62 790

512.01 Yes 3.82 791

524.13 Yes 3.11, 3.58 792

538.12 Yes 4.05 793

502.02 Yes 4.3 794

485.11 Yes 3.62 795

466.1 Yes 3.43 796

460.11 Yes 3.66 797

504.01 Yes 6.43 798

502.02 Yes 4.55 799

493.05 Yes 3.41 800

545.97 Yes 4.41 801

528.08 Yes 3.39 802

482.07 Yes 3.44, 3.71 803

553.11 Yes 3.65 804

493.05 Yes 3.9 805

524.12 Yes 4.35, 6.49 806

498.07 Yes 3.42 807

482.07 Yes 3.99 808

468.06 Yes 3.74 809

502.02 Yes 3.92 810

482.07 Yes 3.68 811

502.02 Yes 4.32 812

550.09 No 813

482.07 Yes 3.84 814

506.12 Yes 3.25 815

482.07 Yes 3.62 816

506.12 Yes 3.57, 4.02 817

572.08 Yes 4.36 818

551.06 Yes 3.92 819

516.03 Yes 820

463.12 Yes 3.69 821

449.11 Yes 3.39 822

435.09 Yes 3.14 823

477.14 Yes 4.18 824

609.06 Tentative 3.91 825

485.11 Yes 3.69 826

485.11 Yes 3.82 827

449.11 Yes 3.37 828

482.07 Yes 3.86 829

463.12 Yes 3.64 830

541.13 No 831

449.11 Yes 3.4 832

463.12 Yes 3.81 833

527.06 No 834

449.11 Yes 3.45 835

485.11 No 836

486.1 No 837

496.11 No 838

464.11 Yes 3.15 839

478.12 Yes 2.76, 3.53,3.93 840

513   Yes 3.75 841

608.14 Yes 3.96 842

518.08 Yes 3.29 843

592.04 Yes 3.8 844

462.13 Yes 3.76, 4.37 845

498.13 Yes 3.99, 4.74 846

516.1 No 847

561.08 No 848

532.09 Tentative 3.92 849

484.09 Yes 3.56 850

532.09 No 851

522.1 No 852

503.01 Yes 3.44 853

550.06 No 854

493.055 Yes 3.3 855

570.07 No 856

524.12 Yes 4.3 857

646.1 Yes 2.96 858

482.07 Yes 3.47 859

580.18 Yes 5.17 860

566.06 Yes 861

560.08 No 862

560.08 Yes 863

536.05 Yes 4.12 864

516.03 Tentative 3.67 865

560.08 Yes 3.33 866

517.03 Yes 3.38, 3.96 867

559.98 Tentative 4.16 868

540.08 No 869

507.07 No 870

556.09 Yes 4.26 871

517.03 No 872

560.08 Yes 3.39 873

593.95 No 874

544.09 Yes 4.21 875

568.11 Yes 4.55 876

496.09 No 877

634   No 878

543.07 Yes 3.59 879

558.1 Yes 3.94 880

584.07 Yes 3.57 881

512.08 No 882

507.07 Yes 3.01 883

583.1 Yes 3.93 884

584.07 Yes 3.5 885

548.08 Tentative 4.47 886

468.06 Yes 3.24 887

486.05 Yes 3.48 888

498.07 Yes 3.29 889

545.97 No 890

558.09 Yes 2.02, 3.17,3.48 891

572.08 Yes 4.07 892

535.98 Yes 4.31 893

519.07 Yes 3.61 894

500.06 Yes 3.46 895

494.07 Yes 3.69 896

537.97 No 897

535.98 No 898

527.01 Yes 3.44 899

579.93 Tentative 4.41 900

562.04 Yes 3.41 901

516.03 Tentative 3.71 902

587.07 Yes 3.68 903

527.01 Yes 3.9 904

558.08 Yes 4.37 905

532.03 Yes 3.46 906

516.03 Tentative 3.99 907

502.02 Yes 3.76 908

535.98 Yes 3.93 909

516.03 Tentative 3.68 910

535.98 Yes 911

584.05 No 912

516.03 Yes 3.84 913

540.08 Yes 3.29 914

516.03 Yes 3.64 915

540.08 Yes 3.58 916

606.05 Yes 4.4 917

585.02 Yes 3.92 918

550   Yes 4.25 919

497.08 Yes 3.67 920

483.07 Yes 3.36 921

469.05 Yes 3.14 922

511.1 Yes 4.05 923

643.02 No 924

519.07 Yes 3.65 925

519.07 Yes 3.83 926

483.07 Yes 3.35 927

516.03 Yes 3.85 928

497.08 Yes 3.59 929

575.09 No 930

459.04 Yes 3.11 931

483.07 Yes 3.4 932

486.08 Yes 2.99 933

497.08 Yes 3.71 934

531.07 Yes 4.27 935

475.01 Yes 3.28 936

561.02 No 937

483.07 Yes 3.42 938

489.03 Yes 3.47 939

505.03 Yes 940

543.01 No 941

519.07 Yes 3.87 942

520.06 No 943

458.05 Yes 2.78 944

530.07 No 945

498.07 Yes 3.2 946

512.08 Yes 3.55 947

546.96 Yes 3.69 948

642.1 Yes 3.98 949

552.04 Yes 3.34 950

626.01 Yes 3.79 951

496.09 Yes 3.77, 4.2 952

532.09 Yes 4, 4.52 953

550.06 Yes 3.74 954

595.05 No 955

566.06 Yes 3.91 956

518.05 Yes 3.59 957

566.06 No 958

474.13 Yes 4 959

460.11 Yes 3.9 960

494.07 Yes 3.9 961

350.17 Tentative 2.46 962

466.2 Yes 3.55, 6.41 963

364.19 Tentative 2.68 964

442.24 Yes 3.55 965

412.15 Tentative 2.78 966

390.21 Tentative 2.86 967

420.25 Tentative 3.55 968

412.19 Tentative 2.88 969

462.21 Yes 3.21 970

378.21 Tentative 2.91 971

424.03 No 972

394.16 Tentative 2.4 973

422.2 Tentative 2.63 974

472.2 Yes 2.15, 3.38 975

474.21 Yes 3.23 976

384.1 Tentative 2.75, 3.26 977

500.2 Yes 3.64, 4.02 978

398.2 Tentative 3.02, 3.66 979

476.2 Yes 4.06 980

446.11 Yes 3.11 981

424.2 Tentative 3.23, 3.88 982

454.2 Yes 4.06, 4.8 983

446.2 Yes 3.22, 3.85 984

496.2 Yes 3.6, 3.71 985

412.2 Tentative 3.28, 3.93 986

457.99 No 987

428.1 Yes 2.68, 3.08 988

456.2 Yes 2.89, 3.34,3.73 989

506.15 Yes 3.84 990

508.17 Yes 3.65 991

356.22 Tentative 2.81 992

472.28 Yes 4.11, 6.46 993

370.24 Tentative 3.05 994

448.28 Yes 4.07 995

418.2 Tentative 3.22 996

396.25 Tentative 3.29 997

426.3 Yes 4.09 998

418.24 Yes 3.29 999

468.25 Yes 3.67 1000

384.25 Tentative 3.35 1001

430.07 No 1002

400.21 Yes 2.72 1003

428.24 Tentative 3.08 1004

478.24 Yes 3.85 1005

480.25 Yes 2.7, 3.73 1006

384.16 Tentative 2.74 1007

474.19 Yes 2.7 1008

468.25 Yes 3.73 1009

402.15 Yes 2.81 1010

502.01 Yes 3.06, 3.9 1011

456.2 Yes 2.15, 3.05 1012

452.15 Yes 3.32 1013

452.15 No 1014

452.15 Yes 2.96 1015

398.17 Tentative 2.8 1016

444.18 Yes 2.76 1017

434.17 Tentative 3.03 1018

402.15 Tentative 2.9 1019

412.19 Yes 2.97 1020

460.19 Yes 3.3 1021

456.18 Yes 3.1 1022

428.18 Tentative 2.88 1023

428.18 Yes 3.01 1024

412.19 Tentative 2.86 1025

409.15 Yes 2.74 1026

520.13 No 1027

398.17 Tentative 2.93 1028

452.08 Yes 3.36 1029

462.07 Yes 3.2 1030

462.07 Yes 3.02 1031

462.07 Tentative 3.21 1032

418.12 Yes 1033

432.1 Yes 3.36, 6.43 1034

418.12 No 1035

452.08 Yes 3.53 1036

440.22 Yes 3.66 1037

452.08 Yes 1038

486.11 No 1039

476.2 Yes 3.21, 3.49,6.51 1040

412.19 Tentative 2.95 1041

444.18 Yes 2.84 1042

428.18 Tentative 2.74 1043

412.19 No 1044

409.2 Yes 2.75, 3.22 1045

418.1 Tentative 3.1, 3.5 1046

508.2 Yes 3.01, 3.39 1047

502.2 Yes 4.18, 4.67 1048

436.1 Yes 3.18, 3.61 1049

490.1 Yes 3.48, 3.99 1050

486.11 Yes 3.79 1051

486.1 Yes 2.79, 3.73,4.25 1052

486.11 Yes 3.33 1053

432.1 Tentative 3.16, 3.62 1054

478.14 Yes 3.09 1055

468.14 Yes 3.42 1056

436.1 Yes 3.27, 3.73 1057

446.2 Yes 3.36, 3.87 1058

494.15 Yes 3.73 1059

490.14 Yes 3.52 1060

462.2 Yes 3.24, 3.75 1061

462.2 Yes 3.43, 4.17 1062

446.2 Yes 3.21, 3.7 1063

443.11 Yes 3.08 1064

554.09 Yes 4.59 1065

432.1 Tentative 3.32, 3.82 1066

486.04 Yes 3.85 1067

496.03 Yes 3.65 1068

496.03 Yes 3.39 1069

496   Yes 3.65, 4.15 1070

452.08 Yes 3.55 1071

466.1 Yes 3.83, 6.53 1072

452.08 Yes 3.37 1073

486.04 No 1074

474.18 Yes 4.14 1075

486.04 Yes 4.01 1076

520.07 Yes 4.15 1077

510.15 Yes 3.96 1078

446.2 Yes 3.31, 3.8 1079

478.1 Yes 3.2, 3.79 1080

462.2 Yes 3.04, 3.49 1081

446.15 No 1082

443.1 Yes 3.07, 3.47 1083

390.21 Tentative 3.1 1084

480.24 Yes 2.71, 3.05 1085

474.3 Yes 4.39 1086

408.2 Tentative 3.19 1087

462.23 Yes 3.5 1088

458.19 Yes 3.77 1089

458.19 Yes 2.7, 3.72 1090

458.19 Yes 3.38 1091

404.22 Tentative 3.21 1092

450.23 Yes 3.12 1093

440.22 Yes 3.48 1094

408.2 Tentative 3.27 1095

418.24 Yes 3.44 1096

466.24 Yes 3.81 1097

462.23 Yes 3.53 1098

434.23 Yes 3.28, 6.53 1099

434.23 Yes 3.45, 6.47 1100

418.24 Tentative 3.33 1101

415.2 Tentative 3.09 1102

526.18 Tentative 2.96 1103

404.22 Tentative 3.35 1104

458.13 Yes 2.7, 3.86 1105

468.12 No 1106

468.12 Yes 3.42 1107

468.12 Yes 3.64 1108

424.17 Yes 3.55 1109

438.18 Yes 3.83 1110

424.17 Yes 3.4 1111

458.13 Yes 4.01 1112

446.27 Yes 4.17 1113

458.13 Yes 2.7, 4.01 1114

492.15 No 1115

482.23 Yes 3.98 1116

418.24 No 1117

450.23 Yes 3.23, 6.51 1118

434.23 Tentative 3.15 1119

418.24 Tentative 3.48 1120

415.2 Yes 3.09 The additional examples below were prepared by individual syntheses. 1121

604.56 yes 1122

570.11 yes 1123

602-59 yes 1124

645.66 yes 1125

590.52 yes 1126

612.52 yes 1127

678.07 yes 1128

587.51 yes 1129

568.53 yes 1130

526.49 yes 1131

526.49 yes 1132

546.91 yes 1133

580.46 yes 1134

546.91 yes 1135

583.59 yes 1136

612.518 yes 1137

630.533 yes 1138

576.485 yes 1139

582.553 yes 1140

587.508 yes 1141

551.475 yes 1142

582.126 yes 1143

632.142 yes 1144

596.109 yes 1145

576.142 yes 1146

582.082 yes 1147

616.527 yes 1148

571.102 yes 1149

521.086 yes 1150

614.555 yes 1151

580.11 yes 1152

565.095 yes 1153

534.081 yes 1154

513.663 yes 1155

517.626 yes 1156

535.069 yes

The compounds listed in Table 1, or their pharmaceutically acceptable salts, may be used in the methods described herein to treat or prevent pain.

It will be understood that when compounds of the present invention contain one or more chiral centers, the compounds of the invention may exist in, and be isolated as, enantiomeric or diastereomeric forms, or as a racemic mixture. The present invention includes any possible enantiomers, diastereomers, racemates or mixtures thereof, of a compound of Formula I. The optically active forms of the compound of the invention may be prepared, for example, by chiral chromatographic separation of a racemate, by synthesis from optically active starting materials or by asymmetric synthesis based on the procedures described thereafter.

It will also be appreciated that certain compounds of the present invention may exist as geometrical isomers, for example E and Z isomers of alkenes. The present invention includes any geometrical isomer of a compound of Formula I. It will further be understood that the present invention encompasses tautomers of the compounds of the formula I.

It will also be understood that certain compounds of the present invention may exist in solvated, for example hydrated, as well as unsolvated forms. It will further be understood that the present invention encompasses all such solvated forms of the compounds of the formula I.

Within the scope of the invention are also salts of the compounds of the formula I. Generally, pharmaceutically acceptable salts of compounds of the present invention may be obtained using standard procedures well known in the art, for example by reacting a sufficiently basic compound, for example an alkyl amine with a suitable acid, for example, HCl or acetic acid, to afford a physiologically acceptable anion. It may also be possible to make a corresponding alkali metal (such as sodium, potassium, or lithium) or an alkaline earth metal (such as a calcium) salt by treating a compound of the present invention having a suitably acidic proton, such as a carboxylic acid or a phenol with one equivalent of an alkali metal or alkaline earth metal hydroxide or alkoxide (such as the ethoxide or methoxide), or a suitably basic organic amine (such as choline or meglumine) in an aqueous medium, followed by conventional purification techniques.

In one embodiment, the compound of formula I above may be converted to a pharmaceutically acceptable salt or solvate thereof, particularly, an acid addition salt such as a hydrochloride, hydrobromide, phosphate, acetate, fumarate, maleate, tartrate, citrate, methanesulphonate or p-toluenesulphonate.

The novel compounds of the present invention are useful in therapy, especially for the treatment of various pain conditions such as chronic pain, neuropathic pain, acute pain, cancer pain, pain caused by rheumatoid arthritis, migraine, visceral pain etc. This list should however not be interpreted as exhaustive.

Compounds of the invention are useful in disease states where degeneration or dysfunction of Bradykinin receptors is present or implicated in that paradigm. This may involve the use of isotopically labeled versions of the compounds of the invention in diagnostic techniques and imaging applications such as positron emission tomography (PET).

Compounds of the invention are useful for the treatment of septic shock, pancreatitis, edema, rhinitis, asthma, colitis, arthritis, hepatorenal syndrome, cancer, (including but not restricted to SCLC, prostrate cancer), bacterial and viral infections, ulcerative colitis, and Alzheimer's Disease.

Compounds of the invention are useful as an analgesic agent for use during general anesthesia and monitored anesthesia care. Combinations of agents with different properties are often used to achieve a balance of effects needed to maintain the anesthetic state (e.g. amnesia, analgesia, muscle relaxation and sedation). Included in this combination are inhaled anesthetics, hypnotics, anxiolytics, neuromuscular blockers and opioids.

Also within the scope of the invention is the use of any of the compounds according to the formula I above, for the manufacture of a medicament for the treatment of any of the conditions discussed above.

A further aspect of the invention is a method for the treatment of a subject suffering from any of the conditions discussed above, whereby an effective amount of a compound according to the formula I above, is administered to a patient in need of such treatment.

Thus, the invention provides a compound of formula I, or pharmaceutically acceptable salt or solvate thereof, as hereinbefore defined for use in therapy.

In a further aspect, the present invention provides the use of a compound of formula I, or a pharmaceutically acceptable salt or solvate thereof, as hereinbefore defined in the manufacture of a medicament for use in therapy.

In the context of the present specification, the term “therapy” also includes “prophylaxis” unless there are specific indications to the contrary. The term “therapeutic” and “therapeutically” should be construed accordingly. The term “therapy” within the context of the present invention further encompasses to administer an effective amount of a compound of the present invention, to mitigate either a pre-existing disease state, acute or chronic, or a recurring condition. This definition also encompasses prophylactic therapies for prevention of recurring conditions and continued therapy for chronic disorders.

The compounds of the present invention are useful in therapy, especially for the therapy of various pain conditions including, but not limited to: acute pain, chronic pain, neuropathic pain, acute pain, back pain, cancer pain, and visceral pain.

In use for therapy in a warm-blooded animal such as a human, the compound of the invention may be administered in the form of a conventional pharmaceutical composition by any route including orally, intramuscularly, subcutaneously, topically, intranasally, intraperitoneally, intrathoracially, intravenously, epidurally, intrathecally, intracerebroventricularly and by injection into the joints.

In one embodiment of the invention, the route of administration may be orally, intravenously or intramuscularly.

The dosage will depend on the route of administration, the severity of the disease, age and weight of the patient and other factors normally considered by the attending physician, when determining the individual regimen and dosage level at the most appropriate for a particular patient.

For preparing pharmaceutical compositions from the compounds of this invention, inert, pharmaceutically acceptable carriers can be either solid or liquid. Solid form preparations include powders, tablets, dispersible granules, capsules, cachets, and suppositories.

A solid carrier can be one or more substances, which may also act as diluents, flavoring agents, solubilizers, lubricants, suspending agents, binders, or table disintegrating agents; it can also be an encapsulating material.

In powders, the carrier is a finely divided solid, which is in a mixture with the finely divided compound of the invention, or the active component. In tablets, the active component is mixed with the carrier having the necessary binding properties in suitable proportions and compacted in the shape and size desired.

For preparing suppository compositions, a low-melting wax such as a mixture of fatty acid glycerides and cocoa butter is first melted and the active ingredient is dispersed therein by, for example, stirring. The molten homogeneous mixture in then poured into convenient sized moulds and allowed to cool and solidify.

Suitable carriers are magnesium carbonate, magnesium stearate, talc, lactose, sugar, pectin, dextrin, starch, tragacanth, methyl cellulose, sodium carboxymethyl cellulose, a low-melting wax, cocoa butter, and the like.

The term composition is also intended to include the formulation of the active component with encapsulating material as a carrier providing a capsule in which the active component (with or without other carriers) is surrounded by a carrier which is thus in association with it. Similarly, cachets are included.

Tablets, powders, cachets, and capsules can be used as solid dosage forms suitable for oral administration.

Liquid form compositions include solutions, suspensions, and emulsions. For example, sterile water or water propylene glycol solutions of the active compounds may be liquid preparations suitable for parenteral administration. Liquid compositions can also be formulated in solution in aqueous polyethylene glycol solution.

Aqueous solutions for oral administration can be prepared by dissolving the active component in water and adding suitable colorants, flavoring agents, stabilizers, and thickening agents as desired. Aqueous suspensions for oral use can be made by dispersing the finely divided active component in water together with a viscous material such as natural synthetic gums, resins, methyl cellulose, sodium carboxymethyl cellulose, and other suspending agents known to the pharmaceutical formulation art.

Depending on the mode of administration, the pharmaceutical composition will preferably include from 0.05% to 99% w (percent by weight), more preferably from 0.10 to 50% w, of the compound of the invention, all percentages by weight being based on total composition.

A therapeutically effective amount for the practice of the present invention may be determined, by the use of known criteria including the age, weight and response of the individual patient, and interpreted within the context of the disease which is being treated or which is being prevented, by one of ordinary skills in the art.

Within the scope of the invention is the use of any compound of formula I as defined above for the manufacture of a medicament.

Also within the scope of the invention is the use of any compound of formula I for the manufacture of a medicament for the therapy of pain.

Additionally provided is the use of any compound according to Formula I for the manufacture of a medicament for the therapy of various pain conditions including, but not limited to: acute pain, chronic pain, neuropathic pain, acute pain, back pain, cancer pain, and visceral pain.

A further aspect of the invention is a method for therapy of a subject suffering from any of the conditions discussed above, whereby an effective amount of a compound according to the formula I above, is administered to a patient in need of such therapy. Additionally, there is provided a pharmaceutical composition comprising a compound of Formula I, or a pharmaceutically acceptable salt thereof, in association with a pharmaceutically acceptable carrier.

Particularly, there is provided a pharmaceutical composition comprising a compound of Formula I, or a pharmaceutically acceptable salt thereof, in association with a pharmaceutically acceptable carrier for therapy, more particularly for therapy of pain. Further, there is provided a pharmaceutical composition comprising a compound of Formula I, or a pharmaceutically acceptable salt thereof, in association with a pharmaceutically acceptable carrier use in any of the conditions discussed above.

In a further aspect, the present invention provides a method of preparing a compound of formula I.

Methods of Preparation

The compounds listed in Table 1 were prepared as single compounds in a combinatorial array. The Table 1, column 4 designation of “yes” indicates that the target mass of the designated compound was found in >50% abundance in the MS spectrum. Similarly, the designation of “tentative” indicates that the target mass of the designated compound was found in 15-50% abundance in the MS spectrum. Likewise the designation “no” indicates that the target mass of the designated compound was found in <15% abundance in MS 'spectrum. It will be understood by those of ordinary skill in the art that a chemical reaction which fails to efficiently yield the desired product within the context of a combinatorial protocol may nonetheless efficiently yield the desired product when the reaction is performed in a single reaction or parallel reaction format, without undue experimentation on the part of the chemist. In this regard, several of the compounds which were not prepared efficiently in the combinatorial array, were subsequently prepared in separate syntheses as shown in the Examples.

The reaction sequence depicted in Scheme 1, infra, describes a process for preparing compounds of formula (I) wherein X is represented by formula (i) or (ii), comprising reacting a compound of general formula II

wherein Y is a protecting group such as CBZ or FMOC, with an alkyl or alkenyl halide, such as allyl bromide in the presence of a base such as cesium carbonate to give compounds of general formula III;

deprotecting the compounds of formula III under standard conditions and then acylated the deprotected products of formula IV with thiophosgene or phosgene to yield isothiocyanates or isocyanates of formula V, respectively:

and subsequently aminating the compounds of formula V combinatorially in a multiwell plate with a selection of different amines to yield compounds of formula I wherein X is represented by formula (i) or (ii) shown above.

The general protocol for the preparation of the combinatorial library is depicted in Scheme 1 and specific experimental details are provided in the Examples below.

-   -   b is 0, 1, or 2; R¹, R², R³, R⁴, and R⁵ are as defined above.

In addition, certain compounds of the present invention and certain intermediates used in the preparation of the compounds of the present invention may be prepared according to one or more of the following general procedures, wherein, unless specified otherwise, b, R¹, R², R³, R⁴ and R⁵ are defined as above.

As illustrated in the scheme above, to a stirred solution of (7-chloro-2,3-dihydro-2-oxo-5-phenyl-1H-1,4-benzodiazepin-3-yl)-, phenylmethyl ester-carbamic acid (10 mmol) in DMF (140 ml) was added cesium carbonate (3.72 g, 11.4 mmol) followed by methyl iodide (2.0 g, 14 mmol). The reaction mixture was stirred at room temperature for 5 hours then concentrated in vacuo. The residue was taken in EtOAc (200 ml) and washed with brine (2×30 ml). The organic phase was then dried over MgSO₄, filtered and concentrated in vacuo. The products were purified by flash chromatography using dichloromethane as the eluent. The products (8 mmol) were added with conc HBr (33% in acetic acid) (50 ml) and were stirred at room temperature for 3 hours. The reaction mixture was poured into ether (300 ml), the precipitate was collected, and then taken in dichloromethane (250 ml) and washed with 2N NaOH (2×50 ml). The organic phase was dried over MgSO4, filtered and concentrated in vacuo to provide the desired compound.

As illustrated in the scheme above, to a solution of 3-aminobenzodiazepines (3 mmol) in dichloroethane (20 ml) and saturated aqueous Na₂CO₃ (20 ml) was added dropwise thiophosgene (0.7 g, 6 mmol). The reaction mixture was stirred at room temperature for 3 hours. The organic phase was separated and the aqueous phase was extracted with dichloromethane (50 ml). The combined organic phases were dried over MgSO₄, filtered and concentrated in vacuo. The product was purified by flash chromatography (100% CH₂Cl₂) to yield a compound of formula V.

As illustrated in the scheme above, to a solution of 4-fluoro-2-methyl-1-nitro-benzene (0.155 g, 1 mmol) in a 1:1 mixture of ethanol and water (20 ml) was added morpholine (0.435 g, 5 mmol). The reaction mixture was heated at 90° C. for 16 h. The solvent was evaporated in vacuo, the residue was taken in dichloromethane (50 ml) and washed with brine (3×10 ml). The organic phase was dried over MgSO₄, filtered, and concentrated in vacuo. The product was taken up in ethanol (20 ml) and heated at reflux. A solution of tin chloride (2M, 2.5 ml) in conc. HCl was added dropwise and heated at reflux for another 30 minutes. The solvent was then evaporated in vacuo, and the residue was treated with a 2M solution of sodium hydroxide until the pH of the solution was >10. The mixture was extracted with dichloromethane (50 ml) and the organic phase was dried over MgSO₄, filtered, and concentrated in vacuo. The product (0.032 g, 0.2 mmol) ((ESI) (M+H)⁺=192) was taken up in dichloroethane (5 ml), and 7-chloro-5-(2-chlorophenyl)-1,3-dihydro-3-isothiocyanato-1-methyl-2H-1,4-benzodiazepin-2-one(0.075 g, 0.2 mmol) was added. The reaction mixture was heated at 70 C for 16 h. The solvent was evaporated in vacuo, and the residue was washed with ether (2×10 ml) to yield the desired compound.

As illustrated in the scheme above, N₂ gas was bubbled through both DME and water for at least 3 hours. A solution of the imidoyl chloride (1 equiv.) in DME (1.5 mL/mmol imidoyl chloride) was placed in a N₂purged flask. Na₂CO₃ (1 equiv.), PdCl₂(dppf) (0.05 equiv.), boronic acid (1 equiv.) and water (0.5 mL/mmol imidoyl chloride) were added sequentially, and the resulting mixture was heated at 100° C. until the imidoyl chloride was consumed (typically 16 h). The reaction was then cooled, diluted with CH₂Cl₂ and water, and the layers were separated. The aqueous phase was extracted with CH₂Cl₂ (3×), and the combined organic phases were dried over Na₂SO₄, filtered, and concentrated in vacuo. The crude product was purified by silica gel column chromatography to provide the desired compound.

General Procedure 5:

As illustrated in the scheme above, boronic acid (1.15 equiv.), Pd₂(dba)₃ (0.015 equiv.), and dry KF (3.3 equiv.) were placed in an oven-dried, N₂ purged flask. A solution of the imidoyl chloride (1 equiv.) in dry THE (2 mL/mmol imidoyl chloride) was added followed by a solution of P(t-Bu)₃ (0.045 equiv., 10% solution in hexanes) in dry THF (1.6 mL/mmol imidoyl chloride). The resulting mixture was heated at reflux until the imidoyl chloride was consumed (typically 16 h). The reaction was then cooled, diluted with EtOAc, and filtered through a small pad of silica gel. The silica was washed well with EtOAc, and the combined organic phases were concentrated in vacuo. The product was purified by silica gel column chromatography to provide the desired compound.

As illustrated in the scheme above, a solution of the benzodiazepine compound (1 equiv.) in dry THF (4 mL/mmol benzodiazepine, or slightly more if solubility was low) was added to a mixture of KHMDS (1.05 equiv., 0.5 M in toluene) and dry THF (2 mL/mmol benzodiazepine) immersed in a −78° C. cooling bath. After stirring for 5 min., a solution of trisyl azide (2.5 equiv) in dry THF (4 mL/mmol benzodiazepine) was added to the reaction, and stirring was continued until all the starting benzodiazepine had been consumed (typically 10 min.). Glacial acetic acid (4.4 equiv) was then added, and the mixture was warmed to 30° C. for 2 h. Saturated NaHCO₃ was added, the layers were separated, and the aqueous phase was extracted with CH₂Cl₂ (4×). The combined organic phases were dried over Na₂SO₄, filtered, and concentrated in vacuo. The product was purified by silica gel column chromatography to provide the corresponding azide compound.

As illustrated in the scheme above, polymer supported triphenylphosphine (Argonaut Technologies, 5-10 equiv) was added to a solution of the azide (1 equiv.) in THF (10 mL/g polymer) and water (0.8 mL/g polymer). The resulting mixture was stirred at room temperature until all of the azide had been consumed (typically overnight). The polymer resin was then removed by filtration, and was washed well with CH₂Cl₂ and MeOH (3× each). The filtrate was concentrated in vacuo, and the residue was redissolved in CH₂Cl₂. Any remaining water was removed with the aid of a separatory funnel, and the organic phase was dried over Na₂SO₄, filtered, and concentrated in vacuo. The amine was purified using a “catch and release” strategy with MP-TsOH resin (Argonaut Technologies): The product was dissolved in CH₂Cl₂ (10 mL/mmol product), and MP-TsOH resin (2.3 equiv) was added. The mixture was stirred for 1 h, and the solvent was removed by filtration and discarded. The resin was rinsed with CH₂Cl₂ and MeOH (3× each), and the washings were discarded as well. The product was then released from the resin by washing with 2M NH₃ in MeOH and CH₂Cl₂ (3× each). Concentration of the filtrate in vacuo provided the corresponding amine compound.

Additional compounds of the present invention may also be prepared according to the methods represented in Schemes 2-4 below, wherein, unless specified otherwise, b, R¹, R², R³, R⁴ and R⁵ are defined as above.

Biological Evaluation I. B2 Bradykinin Binding Assay

A. Human Bradykinin B2 (hB2) Receptor Expression and Membrane Preparation

The cloned human Bradykinin B2 (hB2) receptor in the pCIN vector was purchased from Receptor Biology. The hB2 receptor was stably transfected into HEK 293 S cells and a clonal cell line was generated. Cells were grown in T-flasks with DMEM culture media containing 10% FBS, 2 mM glutamine, 600 μg/ml neomycin and an antibiotic cocktail (100 IU penicillin, 100 μg/ml streptomycin, 0.25 μg/ml amphotericin B). Membranes, expressing the hB2 receptor, were prepared from this cell line according to this protocol: Cells are harvested at 1 to 1.2 million cells/ml, pelleted, and resuspended in ice-cold lysis buffer (50 mM Tris, pH 7.0, 2.5 mM EDTA, with PMSF added just prior to use to 0.5 mM from a 0.5 M stock in DMSO. After lysis on ice for 15 min, the cells are homogenized with a polytron for 10 sec. The suspension is spun at 1000 g for 10 min at 4° C. The supernatant is saved on ice and the pellets resuspended and spun as before. The supernatants from both spins are combined and spun at 46,000 g for 10-30 min. The pellets are resuspended in cold Tris buffer (50 mM Tris/Cl, pH 7.0) at a dilution of 0.2-1 ml per 40 million cells and spun again. The final pellets are resuspended in membrane buffer (50 mM Tris, 0.32 M sucrose, pH 7.0). Aliquots are frozen in dry ice/ethanol and stored at −70° C. until use. The protein concentrations are determined by a modified Lowry with SDS.

B. hB2 Receptor Binding

Membranes expressing the hB2 receptor are thawed at 37° C., passed 3 times through a 25-gauge blunt-end needle, diluted in the bradykinin binding buffer (50 mM Tris, 3 mM MgCl₂, and 1 mg/ml BSA, pH 7.4, 0.02 mg/ml Phenanthroline, 0.25 mg/ml Pefabloc) and 80 μL aliquots containing the appropriate amount of protein (final concentration of 0.25 μg/ml) are distributed in 96-well polystyrene plates (Treff Lab). The IC₅₀ of compounds are evaluated from 10-point dose-response curves, where the serial dilutions are done on a final volume of 150 μL, with 70 μL of ¹²⁵I-Desamino-TyrHOE140 (Kd=0.05) at 50,000 to 60,000 dpm per well (0.03-0.04 nM) in a final volume of 300 μl. The total and non-specific binding are determined in the absence and presence of 0.1 μM (150 μL) of Bradykinin respectively. The plates are vortexed and incubated for 60 minutes at room temperature, filtered through Unifilters-96 GF/B (Canberra Packard), which were presoaked in 0.1% polyethyleneimine, with a harvester using 3 ml of wash buffer (50 mM Tris, pH 7.0, 3 mM MgCl₂). The filters are dried for 1 hour at 55° C. The radioactivity (cpm) is counted in a TopCount (Canberra Packard) after adding 65 μl/well of MS-20 scintillation liquid (Canberra Packard). Compounds of the present invention have demonstrated hB2 receptor binding at concentrations less than 10 μM.

Based on the above assays, the dissociation constant (Ki) for a particular compound of the invention towards a particular receptor is determined using the following equation: Ki=IC ₅₀/(1+[rad]/Kd),

Wherein IC₅₀ is the concentration of the compound of the invention at which 50% displacement has been observed;

[rad] is a standard or reference radioactive ligand concentration at that moment; and

Kd is the dissociation constant of the radioactive ligand towards the particular receptor.

II. GTP[γ]³⁵S Binding Experiments on Bradykinin (B2) Receptors

A. General Information

The procedures below describe how to perform and interpret GTP[γ]³⁵S binding experiments designed to determine the activity of new compounds on the human B2 receptor.

B. General Procedure of the Assay

Human Bradykinin-2 GTP[γ]³⁵S Binding

Human Bradykinin-2 membranes (hB2 293s) are thawed at 37° C., passed 3 times through a 25-gauge blunt-end needle and diluted in the GTPγS binding buffer for the assay (50 mM Hepes, pH 7.4; 200 mM NaCl; 1 mM EDTA; 5 mM MgCl₂. To this added freshly prepared 1 mM DTT, 0.5% BSA, 1 μM GDP. The EC50 and Emax of compounds are evaluated from 10-point dose-response curves done in 300 μl with the appropriate amount of membrane protein and 100,000-120,000 dpm of GTPγ³⁵S per well (0.11-0.14 nM). Bradykinin (1-9) is used as the standard agonist at hB2. The ranges of concentrations tested should include a maximal concentration of 0.1 μM bradykinin in order to establish the E_(max).

The plates are vortexed and incubated for 60 minutes at room temperature, filtered on GF/B Unifilters (presoaked in water) with the Packard harvester using 4 ml/well of wash buffer (50 mM Tris, 5 mM MgCl₂, 50 mM NaCl, pH 7.0), minimum. The filters are dried for 1 hour at 55° C. The radioactivity (cpm) is counted in a TopCount (Packard) after adding 65 μl/well of MS-20 scintillation liquid.

Antagonist reversal studies are done in a similar manner except that the compound dose-response curve's are performed in the presence of a constant concentration of agonist (approx. 80% bradykinin E_(max); ˜5 nM). A standard B2 Antagonist is used as the reference antagonist at hB2. The ranges of antagonist concentrations tested should include a maximal concentration of 3 μM of the standard B2 Antagonist in order to establish the maximal displacement (D_(max)).

C. Radioligand: Preparation of GTP[γ]³⁵S

GTP[γ]³⁵S is acquired from Perkin-Elmer (250 μCi/20 μl). It is diluted from with 10 mM DTT, 50 mM Tris, pH 7 (dilute in 2 ml, 1.0 mCI/20 μ). Sonicate the solution, filter through a 0.45 μm filter, and freeze aliquots at −70° C. For the experiment, use ˜0.3 nM dilution of this tracer in the GTP binding buffer.

D. Data Analysis

The EC₅₀ and E_(max) of compounds are evaluated from 10-point dose-response curves done in 300 μl with the appropriate amount of membrane protein and GTPγ³⁵S per well and are calculated in Activity base with ExcelFit. The basal and maximal stimulated binding are determined in the absence and presence of standard reference compounds, respectively.

The stimulation (Stim) in the presence of compounds is expressed as the percentage of D_(max) of the reference antagonist. Values of IC₅₀, Ki′ and D_(max) for ligands capable of competing for agonist stimulated binding are calculated in Activity Base. Mean±S.E.M. values of IC₅₀, Ki′ and % D_(max) are reported for ligands tested in at least three dose-response curves.

Biological data for particular testing samples (as listed in Table 2) of the compounds of the invention are listed in Table 3 below.

TABLE 2 List of the test samples used in the Biological Evaluation Test Sample Nos: Structure of the Test Sample 1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

TABLE 3 Biological Data for the testing samples as listed in Table 2 Test Sample Nos. Ki (hB2) (nM) 1-33 43-3110

EXAMPLES

The invention will further be described in more detail by the following Examples which describe methods whereby compounds of the present invention may be prepared, purified, analyzed and biologically tested, and which are not to be construed as limiting the invention.

Intermediate 1 3-amino-7-chloro-1,3-dihydro-1-methyl-5-phenyl-2H-1,4-benzodiazepin-2-one

Following General Procedure 1, INTERMEDIATE 1 was obtained as pale brown solid (2.5 g, 77%) and used for the subsequent reaction without further purification. MS (ESI) (M+H)⁺=300.

Intermediate 2 3-amino-5-(2-bromophenyl)-7-fluoro-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one

Following General procedure 1, INTERMEDIATE 2 was obtained as a thick pale brown oil (0.5 g, 17%) and used for the subsequent reaction without further purification.

Intermediate 3 3-amino-5-cyclohexyl-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one

Following General Procedure 1, after flash chromatography (100% EtOAc), INTERMEDIATE 3 was obtained as a pale brown solid (1.25 g, 45%). MS (ESI) (M+H)⁺=272

Intermediate 4 3-amino-7-chloro-5-(2-chlorophenyl)1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one

Following General Procedure 1, INTERMEDIATE 4 was obtained as a thick pale brown oil (1.8 g, 65%) and used for the subsequent reaction without further purification. MS (ESI) (M+H)⁺=334

Intermediate 5 3-amino-1,3-dihydro-1-methyl-5-phenyl-2H-1,4-benzodiazepin-2-one

To a stirred solution of (2,3-dihydro-2-oxo-5-phenyl-1H-1,4-benzodiazepin-3-yl)-, phenylmethyl ester-carbamic acid, (4.0 g, 10.3 mmol) in toluene (100 ml) was added Aliquat336 (1.0 g) and 50% aqueous sodium hydroxide (20 ml) followed by methyl iodide (5.0 g, 35 mmol). The reaction mixture was stirred at room temperature for 17 hours. The solvent was evaporated in vacuo and the residue was taken in dichloromethane (150 ml). The organic phase was washed with 2N sodium hydroxide (50 ml) and brine (50 ml); the organic phase was dried over MgSO₄, filtered and concentrated in vacuo. The residue was triturated with hexane and the precipitate was treated with conc. HBr as in General Procedure 1. INTERMEDIATE 5 was obtained as a pale brown solid (1.63 g, 59%) and used for the subsequent reaction without further purification. MS (ESI) (M+H)⁺=266

Intermediate 6 3-amino-7-chloro-1,3-dihydro-5-phenyl-1-(2-propenyl)-2H-1,4-benzodiazepin-2-one

To a stirred solution of (7-chloro-2,3-dihydro-2-oxo-5-phenyl-1-H-1,4-benzodiazepin-3-yl)-, phenylmethyl ester-carbamic acid (1 mmol) in DMF (10 ml) was added cesium carbonate (0.370 g, 1.14 mmol) followed by allyl bromide (0.134 g, 1.1 mmol). The reaction mixture was stirred at room temperature for 5 hours. The solvent was evaporated in vacuo and the residue was taken in EtOAc (50 ml). The organic phase was washed with brine (2×10 ml); the organic phase was dried over MgSO₄, filtered and concentrated in vacuo. The crude products were purified by flash chromatography using dichloromethane as the eluent. The crude product was added to conc. HBr (33% in acetic acid) (10 ml) and stirred at room temperature for 3 hours. The reaction mixture was poured into ether (100 ml), the precipitate was collected and then taken in dichloromethane (50 ml) and washed with 2N NaOH (2×10 ml). The organic phase dried over MgSO4, filtered and concentrated in vacuo. The title compound was obtained as a pale yellow solid (0.19 g, 60%) and used for the subsequent reaction without further purification. MS (ESI) (M+H)⁺=326

Intermediate 7 7-chloro-1,3-dihydro-3-isothiocyanato-1-methyl-5-phenyl-2H-1,4-benzodiazepin-2-one

Following General Procedure 2, INTERMEDIATE 7 was obtained as a pale yellow solid (0.7 g, 69%). ¹H-NMR (CDCl₃): δ 7.66-7.64 (m, 2H), 7.60-7.57 (dd, J=2.4 Hz and 8.8 Hz, 1H), 7.55-7.51 (m, 1H), 7.47-7.43 (m, 2H), 7.36-7.34 (m, 2H) and 3.48 (s, 4H) MS (ESI) (M+H)⁺=342

Intermediate 8 7-chloro-5-(2-chlorophenyl)-1,3-dihydro-3-isothiocyanato-1-methyl-2H-1,4-benzodiazepin-2-one

Following General Procedure 2, after flash chromatography (dichloromethane), the title compound was obtained as pate yellow solid (2.2 g, 65%). ¹H-NMR (CDCl₃): δ 7.66-7.64 (m, 1H), 7.56-7.53 (dd, J=2.4 Hz and 8.8 Hz, 1H), 7.46-7.43 (m, 2H), 7.39-7.37 (m, 1H), 7.05 (d, J=2.0 Hz, 1H), 3.73 (s, 1H) and 3.51 (s, 3H). MS (ESI) (M+H)⁺=376

Example 1 N-(7-chloro-2,3-dihydro-1-methyl-2-oxo-5-phenyl-1H-1,4-benzodiazepin-3-yl)-N′-(5isoquinolinyl)-thiourea

As illustrated in the scheme above, to a solution of 7-chloro-1,3-dihydro-3-isothiocyanato-1-methyl-5-phenyl-2H-1,4-benzodiazepin-2-one (INTERMEDIATE 7) (0.035 g, 0.1 mmol) in dichloromethane (5 ml) was added 5-isoquinolinamine (0.015 g, 0.1 mmol). The reaction mixture was heated at reflux for 17 hours. The solvent was evaporated in vacuo, the residue was triturated with ether and the title compound was obtained as a colorless solid (20 mg, 40%). ¹H-NMR (CDCl₃): δ 9.33 and 9.19 (2×s, 1H), 8.63 and 8.49 (2×d, J=5.6 Hz and J=6.0 Hz, 1H), 8.34 (br s, 1H), 8.02 (d, J=8.4 Hz, 1H), 7.92 (t, J=11.6 Hz, 2H), 7.58-7.32 (m, 10H), 6.03 (d, J=7.6 Hz, 1H) and 3.3 (s, 3H). MS (ESI) (M+H)⁺=486

Example 2 N-(7-chloro-2,3-dihydro-1-methyl-2-oxo-5-phenyl-1H-1,4-benzodiazepin-3-yl)-N′-[4(dimethylamino)phenyl]-thiourea

To a solution of 7-chloro-1,3-dihydro-3-isothiocyanato-1-methyl-5-phenyl-2H-1,4-benzodiazepin-2-one (INTERMEDIATE 7) (0.342 g, 1 mmol) in dichloromethane (20 ml) was added N,N-dimethyl-1,4-benzenediamine (0.136 g, 1 mmol). The reaction mixture was heated at reflux for 17 hours. The solvent was evaporated in vacuo, the residue was triturated with ether and the title compound was obtained as a colorless solid (0.167 g, 35%). ¹H-NMR (CDCl₃): δ 7.70-7.22 (m, 10H), 6.75 (d, J=8.8 Hz, 2H), 6.05 (d, J=7.6 Hz, 1H), 3.42 (s, 3H) and 2.98 (s, 6H). MS (ESI) (M+H)⁺=478.

Example 3 N-(7-chloro-2,3-dihydro-1-methyl-2-oxo-5-phenyl-1H-1,4-benzodiazepin-3-yl)-N′-[4(dimethylamino)-1-naphthalenyl]-N-methyl-thiourea

To a stirred solution of carboxybenzyl-3-amino-7-chloro-5-phenyl-2-oxo-1,4-benzodiazepine (EXAMPLE 6) (1 mmol) in DMF(10 ml) was added cesium carbonate (5 mmol) followed by methyl iodide (2.2 mmol). The reaction mixture was stirred at room temperature for 5 hours. The solvent was evaporated in vacuo and the residue was taken in EtOAc (50 ml). The organic phase was washed with brine (2×10 ml); the organic phase was dried over MgSO₄, filtered and concentrated in vacuo. The product (0.8 mmol) was added to conc. HBr (35% in acetic acid) (10 ml) and was stirred at room temperature for 3 hours. The reaction mixture was poured into ether (30 ml), the precipitate was collected, and taken in dichloromethane (25 ml) and washed with 2N NaOH (2×10 ml). The organic phase was dried over MgSO₄, filtered and concentrated in vacuo. The product was used for the subsequent steps without further purification.

To a solution of the product above (0.03 g, 1 mmol) in dichloroethane (5 ml) was added 4-isothiocyanato-N,N-dimethyl-1-naphthalenamine (0.025 g, 1 mmol). The reaction mixture was stirred at 70 C for 4 hours. The solvent was evaporated in vacuo, the residue was triturated with ether and the title compound was obtained as a colorless solid (28 mg, 52%). ¹H-NMR (CDCl₃): δ 8.24 (m, 1H), 7.92 (m, 1H), 7.67 (d, J=7.2 Hz, 2H), 7.52-7.40 (m, 8H), 7.31 (d, J=1.6 Hz, 1H), 7.05 (d, J=8.0 Hz, 2H), 3.78 (s, 3H), 3.41 (s, 3H) and 2.91 (s, 6H). MS (ESI) (M+H)⁺=542.

Example 4 N-[7-chloro-5-(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzodiazepin-3-yl]-N′-[4-(dimethylamino)-1-naphthalenyl]-thiourea

To a solution of 3-amino-7-chloro-5-(2-chlorophenyl)-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one (INTERMEDIATE 8) (0.03 g, 1 mmol) in dichloroethane (5 ml) was added 4-isothiocyanato-N,N-dimethyl-1-naphthalenamine (0.025 g, 1 mmol). The reaction mixture was stirred at 70° C. for 4 hours. The solvent was evaporated in vacuo, the residue was triturated with ether and the title compound was obtained as a colorless solid (35 mg, 60%). ¹H-NMR (CDCl₃): δ 8.26 (d, J=8.4 Hz, 1H), 8.03 (d, J=7.6 Hz, 1H), 7.93 (br s, 1H), 7.73-7.06 (m, 11H) 6.10 (d, J=7.6 Hz, 1H), 3.40 (s, 3H) and 2.92 (s, 6H). MS (ESI) (M+H)⁺=562.

Example 5 N-[7-chloro-2,3-dihydro-2-oxo-5-phenyl-1-(2-propenyl)-1H-1,4-benzodiazepin-3-yl]-N′-[4-(dimethylamino)-1-naphthalenyl]-thiourea

A solution of 3-amino-7-chloro-1,3-dihydro-5-phenyl-1-(2-propenyl)-2H-1,4-benzodiazepin-2-one (INTERMEDIATE 6) (0.162 g, 0.5 mmol) in dichloroethane (5 ml) was added 4-isothiocyanato-N,N-dimethyl-1-naphthalenamine (0.115 g, 0.5 mmol). The reaction mixture was stirred at 70° C. for 4 hours; The solvent was evaporated in vacuo, the residue was triturated with ether and the title compound was obtained as a colorless, solid (0.152 g, 55%). ¹H-NMR (CDCl₃): δ 8.29-8.27 (m, 1H), 8.08-8.06 (m, 1H), 7.90 (s, 1H), 7.60-7.36 (m, 10H), 7.32 (d, J=2.4 Hz, 1H), 7.09 (d, J=8 Hz, 1H), 6.14 (d, J=7.6 Hz, 1H), 5.76-5.69 (m, 1H), 5.15 (s, 1H), 5.12 (dd, J=1.2 and 7.0 Hz, 1H), 4.57-4.39 (m, 2H) and 2.93 (s, 6H). MS (ESI) (M+H)⁺=554.

Example 6 N-(7-chloro-2,3-dihydro-1-methyl-2-oxo-5-phenyl-1H-1,4-benzodiazepin-3-yl)-N′-[4-(dimethylamino)-1-naphthalenyl]-thiourea

To a solution of 3-amino-7-chloro-5-phenyl-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one (INTERMEDIATE 1) (0.299 g, 1 mmol) in dichloroethane (15 mi) was added 4-isothiocyanato-N,N-dimethyl-1-naphthalenamine (0.230 g, 1 mmol). The reaction mixture was stirred at room temperature for 17 hours. The solvent was evaporated in vacuo, the residue was triturated with ether and the title compound was obtained as a colorless solid (0.345 g, 65%). ¹H-NMR (CD₃OD): δ 8.25 (d, 8.4 Hz, 2H), 8.00 (d, J=8.4 Hz, 1H), 7.88 (d, J=8.4 Hz, 1H), 7.85-7.81 (m, 2H), 7.77-7.72 (m, 2H), 7.65 (d, J=8.8 Hz, 1H), 7.61-7.58 (m, 4H), 7.51-7.47 (m, 2H), 7.29 (d, J=2.4 Hz, 1H), 6.01 (s, 1H) and 3.49 (s, 9H). MS (ESI) (M+H)⁺=528.

Example 7 N-(7-chloro-2,3-dihydro-1-methyl-2-oxo-5-phenyl-1H-1,4-benzodiazepin-3-yl)-N′-(4-methoxy-2-methylphenyl)-urea

To a solution of 3-amino-7-chloro-5-phenyl-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one (INTERMEDIATE 1) (0.299 g, 1 mmol) in dichloromethane (15 ml) was added 1-isocyanato-4-methoxy-2-methyl-benzene (0.163 g, 1 mmol). The reaction mixture was stirred at room temperature for 17 hours. The solvent was evaporated in vacuo, the residue was triturated with ether and the title compound was obtained as a colorless solid (0.197 g, 42%). ¹H-NMR (CDCl₃): δ 7.57-7.48 (m, 3H), 7.47-7.44 (m, 1H), 7.39-7.38 (m, 3H), 7.36-7.29 (m, 2H), 6.77-6.73 (m, 2H), 6.71 (d, J=2.8 Hz, 1H), 6.61 (s, 1H), 5.51 (d, J=8.4 Hz, 1H), 3.78 (s, 3H), 3.39 (s, 3H) and 2.29 (s, 3H). MS (ESI) (M+H)⁺=463.

Example 8 N-(7-chloro-2,3-dihydro-1-methyl-2-oxo-5-phenyl-1H-1,4-benzodiazepin-3-yl)-N′-[4-(dimethylamino)-1-naphthalenyl]-urea

As illustrated in the scheme above, a mixture of 4-(dimethylamino)-1-naphthalenecarboxylic acid (0.1 mmol), diphenylphosphoryl azide (0.15 mmol) and triethylamine (0.3 mmol) in toluene (10 ml) was heated at reflux overnight. The solvent was evaporated in vacuo, and then the residue was redissolved in dichloroethane, added to INTERMEDIATE 1 (0.08 mmol) and heated at 70° C. for 4 hours. The solvent was evaporated in vacuo and the residue was taken in dichloromethane (50 ml). The organic phase was washed with brine (2×10 ml); the organic phase was dried over MgSO₄, filtered and concentrated in vacuo. The residue was triturated with ether and the title compound was obtained as a colorless solid (9 mg, 22%); ¹H-NMR (CDCl₃): δ 8.27 (m, 1H), 8.11 (m, 1H), 7.60 (d, J=8 Hz, 1H), 7.77-7.29 (m, 10H), 7.08 (d, J=8.4 Hz, 1H), 6.75 (br s, 2H), 5.56 (d, J=8.4 Hz, 1H), 3.39 (s, 3H) and 2.90 (s, 6H); MS (ESI) (M+H)⁺=512.

Example 9 N-[5-(2-bromophenyl)-7-fluoro-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzo-diazepin-3-yl]-N′-[4-(dimethylamino)-1-naphthalenyl]-thiourea

As illustrated in the scheme above, INTERMEDIATE 2 (0.018 g, 0.05 mmol) and 4-isothiocyanato-N,N-dimethyl-1-naphthalenamine (0.011 g, 0.05 mmol) was heated in dichloroethane (4 mL) at 70° C. overnight. The solvent was evaporated in vacuo and the residue was trituated with ether (2×10 ml). The title compound (0.021 g, 72%) was obtained as a colorless solid. ¹H-NMR (CDCl₃): δ 8.26 (m, 1H), 8.06 (m, 1H), 7.85 (s, 1H), 7.79 (m, 1H), 7.69-7.24 (m, 8H), 7.08 (d, J=8.0 Hz, 1H), 7.02 (t, J=8.4 Hz, 1H), 6.06 (d, J=8.0 Hz, 1H), 3.39 (s, 3H), 2.93 (s, 6H). (ESI) (M+H)⁺=591.

Example 10 N-[7-chloro-5-(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzo-diazepin-3-yl]-N″-cyano-N′-[4-(4-morpholinyl)-1-naphthalenyl]-guanidine (E and Z isomers Separated but not Identified)

As illustrated in the scheme above, a solution of N-[7-chloro-5-(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzodiazepin-3-yl]-N′-[4-(4-morpholinyl)-1-naphthalenyl]-thiourea (EXAMPLE 11, made according to General Procedure 3) (0.058 g, 0.1 mmol) and silver triflate (0.077 g, 0.3 mmol) in dichloromethane (2 ml) was combined with cyanamide disodium salt (1 mmol) and stirred at room temperature for 3 h. The reaction mixture was then diluted with dichloromethane (10 ml) and washed with brine (2×5 ml). The organic phase was dried over MgSO₄, filtered and concentrated in vacuo. The residue was washed with ether (2×10 ml) and the polar isomer of the title compound (0.023 g, 380%) was obtained as pale yellow solid. The ether layer was concentrated in vacuo and purified by column chromatography (1:1 EtOAc:CH₂Cl₂) to give the non-polar isomer of the title compound (0.009 g, 15%) as a colorless solid.

Example 10A N-[7-chloro-5-(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzo-diazepin-3-yl]-N″-cyano-N′-[4-(4-morpholinyl)-1-naphthalenyl]-guanidine (polar isomer)

¹H-NMR (CDCl₃): δ 8.26 (m, 1H), 7.9 (m, 2H), 7.69 (d, J=12.0 Hz, 2H), 7.53-7.46 (m, 4H), 7.39 (m, 1H), 7.30-7.23 (m, 4H), 7.11 (m, 2H), 3.98 (t, J=9.2 Hz, 2H), 3.48 (s, 3H) and 3.12 (br.s, 6H). (ESI) (M+H)⁺=612

Example 10B N-[7-chloro-5(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzo-diazepin-3-yl]-N″-cyano-N′-[4-(4-morpholinyl)-1-naphthalenyl]-guanidine (nonpolar isomer)

¹H-NMR (CDCl₃): δ 8.25 (m, 1H), 8.03 (m, 1H), 7.66-7.53 (m, 5H), 7.44-7.40 (m, 3H), 7.39-7.30 (m, 2H), 7.09 (m, 2H), 6.54 (d, J=7.6 Hz, 1H), 5.48 (d, J=7.6 Hz, 1H), 3.99 (t, J=9.2 Hz, 4H), 3.39 (s, 3H) and 3.15 (br.s, 4H). (ESI) (M+H)⁺=612.

Example 11 N-[7-chloro-5-(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzodiazepin-3-yl]-N′-[4-(4-morpholinyl)-1-naphthalenyl]-thiourea

Following General Procedure 3, the title compound (0.078 g, 65%) was obtained as a white solid. ¹H-NMR (CDCl₃): δ 8.4 (m, 1H), 8.08 (m, 2H), 7.72 (m, 1H), 7.64 (d, J=8 Hz, 1H), 7.6-7.5 (m, 4H), 7.39 (t, J=4.4 Hz, 2H), 7.30 (d, J=8.8 Hz, 2H), 7.10 (m, 2H), 6.09 (d, J=7.6 Hz, 1H), 3.98 (t, J=4.4 Hz, 4H) 3.40 (s, 3H), and 3.13 (m, 4H). (ESI) (M+H)⁺=604.

Example 12 N-[7-chloro-5-(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzo-diazepin-3-yl]-N′-[4(4-morpholinyl)-1-naphthalenyl]-guanidine

As illustrated in the scheme above, to a solution of N-[7-chloro-5-(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzodiazepin-3-yl]-N′-[4-(4-morpholinyl)-1-naphthalenyl]-thiourea (EXAMPLE 11) (0.029 g, 0.05 mmol) and silver triflate (0.038 g, 0.15 mmol) in dichloromethane (2 ml) was added a solution of ammonia (0.25 ml, 2M in methanol). The reaction mixture was stirred at room temperature for 3 h, diluted with dichloromethane (10 ml) and washed with brine (2×5 ml). The organic layer was dried with MgSO4 filtered, concentrated in vacuo. The residue was triturated with ether to give the title compound (0.008 g, 28%) as a pale yellow solid. ¹H-NMR (CDCl₃): δ 8.22 (d, J=9.2 Hz, 1H), 8.11 (d, J=8.4 Hz, 1H), 7.75 (br.s, 1H), 7.50-7.41 (m, 7H), 7.31 (d, J=8.8 Hz, 1H), 7.04 (m, 2H), 5.8 (br.s, 1H), 3.96 (t, J=9.2 Hz, 4H) 3.51 (s, 3H) and 3.05 (br.s, 4H).). (ESI) (M+H)⁺=587.

Example 13 N-[7-chloro-5-(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzo-diazepin-3-yl]-N′-[2-methyl-4-(4-morpholinyl)phenyl]-thiourea

Following General Procedure 3, the title compound (0.096 g, 85%) was obtained as a colorless solid. ¹H-NMR (CDCl₃): δ 7.74 (m, 1H), 7.54 (s, 1H), 7.51 (m, 1H), 7.42-7.39 (m, 3H), 7.34-7.31 (m, 2H), 7.24 (d, J=8.8 Hz, 1H), 7.09 (d, J=2.4 Hz, 1H), 6.80-6.76 (m, 2H), 6.09 (d, J=7.6 Hz, 1H), 3.85 (t, J=9.6 Hz, 4H), 3.44 (s, 3H), 3.17 (t, J=10.0 Hz, 4H) and 2.32 (s, 3H). (ESI) (M+H)⁺=568.

Example 14 N-[7-chloro-5-(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzo-diazepin-3-yl]-N′-[4-(dimethylamino)-2-methylphenyl]-thiourea

Following the General Procedure 3, the title compound (0.052 g, 51%) was obtained as a pale brown solid. ¹H-NMR (CDCl₃): δ 7.75 (m, 1H), 7.52 (dd, J=8.8 Hz, 1H), 7.47 (s, 1H), 7.42-7.40 (m, 3H), 7.34-7.31 (m, 2H), 7.19 (d, J=8.0 Hz, 1H), 7.09 (d, J=2.0 Hz, 1H), 6.60 (br.s, 1H), 6.58 (br.s, 1H), 6.09 (d, J=8.0 Hz, 1H), 3.44 (s, 3H), 2.97 (s, 6H) and 2.30 (s, 3H). (ESI) (M+H)+=526.

Example 15 N-[7-chloro-5-(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzo-diazepin-3-yl]-N′-[4-(dimethylamino)-3-methylphenyl]-thiourea

Following General Procedure 3, the title compound (0.034 g, 35%) was obtained as a pale brown solid. ¹H-NMR (CDCl₃): δ 7.86 (d, J=7.6 Hz, 1H), 7.76 (m, 1H), 7.53 (dd, J=8.8 Hz, 1H), 7.42-7.39 (m, 2H), 7.36-7.32 (m, 2H), 7.17 (dd, J=8.8 Hz, 1H), 7.12 (d, J=2.0 Hz, 1H), 7.10 (d, J=2.4 Hz, 1H), 7.06 (d, J=8.4 Hz, 1H), 6.09 (d, J=7.6 Hz, 1H), 3.46 (s, 3H), 2.71 (s, 6H) and 2.33 (s, 3H). (ESI) (M+H)⁺=526.

Example 16 N-[7-chloro-5-(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzo-diazepin-3-yl]-N′-[3-chloro-4-dimethylamino)phenyl]-thiourea

Following the General Procedure 3, the title compound (0.025 g, 23%) was obtained as a pale brown solid. ¹H-NMR (CDCl₃): δ 7.86 (s, 1H), 7.83 (d, J=7.2 Hz, 1H), 7.5 (m, 1H), 7.53 (dd, J=8.8 Hz, 1H), 7.43-7.40 (m, 2H), 7.35-7.33 (m, 2H), 7.28 (m, 1H), 7.10 (s, J=2.0 Hz, 1H), 7.05 (d, J=7.2 Hz, 1H), 6.06 (d, J=7.2 Hz, 1H), 3.47 (s, 3H) and 2.82 (s, 6H). (ESI) (M+H)⁺=546.

Example 17 N-[7-chloro-5-(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzo-diazepin-3-yl]-N′-[4-(dimethylamino)-3-(trifluoromethyl)phenyl]-thiourea

Following General Procedure 3, the title compound (0.017 g, 18%) was obtained as a pale brown solid. ¹H-NMR (CDCl₃): δ 7.93 (s, 1H), 7.85 (d, J=7.2 Hz, 1H), 7.38 (m, 1H), 7.58 (d, J=8.4 Hz, 1H), 7.55-7.53 (m, 2H), 7.42-7.40 (m, 2H), 7.34 (m, 3H), 7.10 (s, J=2.0 Hz, 1H), 6.06 (d, J=7.6 Hz, 1H), 3.47 (s, 3H) and 2.76 (s, 6H). (ESI) (M+H)⁺=580.

Example 18 N-[7-chloro-5(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzo-diazepin-3-yl]-N′-[4-chloro-2-(dimethylamino)phenyl]-thiourea

Following the General Procedure 3, the title compound (0.024 g, 22%) was obtained as a pale brown solid. ¹H-NMR (CDCl₃): δ 8.55 (d, J=7.2 Hz, 1H), 7.84 (s, 1H), 7.77 (t, J=4.8 Hz, 1H), 7.53 (dd, J=8.8 Hz, 1H), 7.46 (d, J=8.4 Hz, 1H), 7.42-7.41 (m, 2H), 7.36-7.34 (m, 3H), 7.11-7.02 (m, 2H), 60.6 (d, J=7.2 Hz, 1H), 3.48 (s, 3H) and 2.75 (s, 6H). (ESI) (M+H)⁺=546.

Example 19 N-[7-chloro-5-(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzo-diazepin-3-yl]-N′-[4-(diethylamino)-2-(dimethylamino)phenyl]-thiourea

Following the General Procedure 3, the title compound (0.040 g, 35%) was obtained as a pale gray solid. ¹H-NMR (CDCl₃): δ 9.4 (s, 1H), 8.22 (s, 1H), 7.76 (m, 1H), 7.52 (dd, J=8.8 Hz, 1H), 7.40 (m, 2H), 7.35 (m, 2H), 7.10 (d, J=2.4 Hz, 1H), 7.03 (d, J=8.8 Hz, 1H), 6.84 (s, 1H), 6.49 (dd, J=8.8 Hz, 1H), 6.11 (d, J=7.2 Hz, 1H), 3.47 (s, 3H) 3.33 (m, 4H), 2.65 (s, 6H) and 1.38 (t, J=14.0 Hz, 6H). ESI) (M+H)⁺=583.

Example 20 N-[(1E)-[[7-Chloro-5-(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzodiazepin-3-yl]amino][[4-(4-morpholinyl)-1-naphthalenyl]amino]methylene]urea

A mixture of N-[7-chloro-5-(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzodiazepin-3-yl]-N″-cyano-N′-[4-(4-morpholinyl)-1-naphthalenyl]guanidine (EXAMPLE 10) (21.9 mg, 35.8 μmol), water (4.6 μL, 260 μmol) and trifluoroacetic acid (19.8 μL, 257 μmol) in THF (3 mL) was heated to reflux for 43 h. The reaction was concentrated in vacuo, and the residue was purified by reverse phase HPLC (gradient 20-70% CH₃CN in H₂O) to provide the title compound (0.0092 g, 35%) as its TFA salt. ¹H-NMR (CD₃OD): δ 8.37 (br s, 1H), 8.06 (br s, 1H), 7.75-7.41 (br m, 9H), 7.26 (br s, 1H), 7.04 (br s, 1H), 5.66 (br s, 1H), 4.00 (br s, 4H), 3.58 (br s, 3H), 3.17 (br s, 4H). HRMS calculated for (C₃₂H₂₉Cl₂N₇O₃+H) (M+H)⁺: 630.1787. Found (ESI): 630.1800.

Example 21 N′-[7-Chloro-5-(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzodiazepin-3-yl]-N-methyl-N-[2-methyl-4-(4-morpholinyl)phenyl]thiourea

A solution of 2-methyl4-(4-morpholinyl)benzenamine (53.3 mg, 0.277 mmol) and diisopropylethylamine (0.063 mL, 0.36 mmol) in CH₂Cl₂ (1 mL) was cooled to 0° C. Methyl chloroformate (0.024 mL, 0.31 mmol) was added dropwise, and then the reaction was allowed to warm to room temperature and stir overnight. The reaction was diluted with CH₂Cl₂ (20 mL) and washed with brine (10 mL). The organic phase was dried over Na₂SO₄, filtered, and concentrated in vacuo. The crude product was then suspended in a 1:2 mixture of Et₂O:THF (6 mL). A solution of LiAlH₄ in Et₂O (0.34 mL of a 1 M solution, 0.34 mmol) was added dropwise, and then the reaction mixture was heated to reflux for 1.5 h. The reaction was cooled, diluted with additional Et₂O (8 mL), and quenched with Na₂SO₄5H₂O (0.98 g, 4.2 mmol). After stirring for 15 minutes, the mixture was filtered and the reaction was concentrated in vacuo. A portion of this crude aniline (0.0580 g, 0.281 mmol) was dissolved in (CH₂Cl)₂ (8 mL), and 7-chloro-5-(2-chlorophenyl)-1,3-dihydro-3-isothiocyanato-1-methyl-2H-1,4-benzodiazepin-2-one (0.106 g, 0.281 mmol) was added. The resulting mixture was heated at 70° C. for 14 h. The reaction was cooled and concentrated in vacuo, and the residue was purified by silica gel column chromatography (7:1 CH₂Cl₂:EtOAc) to provide the title compound (0.1211 g, 74%). Due to hindered rotation about one of the bonds, rotamers were observed in the ¹H-NMR spectrum. ¹H-NMR (CDCl₃): δ 7.77-7.68 (m, 1H), 7.51 (dd, J=2.4 Hz, J=8.8 Hz, 1H), 7.43-7.36 (m, 2H), 7.34-7.28 (m, 2H), 7.16 (t, J=8.4 Hz, 1H), 7.10-7.01 (m, 2H), 6.86-6.76 (m, 2H), 6.14 and 6.10 (2×d, J=8.0 Hz, J=7.6 Hz, 1H), 3.88-3.82 (br m, 4H), 3.60 and 3.59 (2×s, 3H), 3.41 (s, 3H), 3.19 (br s, 4H), 2.24 and 2.23 (2×s, 3H). MS (ESI) (M+H)⁺=582. HRMS calculated for (C₂₉H₂₉Cl₂N₅O₂S+H) (M+H)⁺: 582.1497. Found (ESI): 582.1448.

Intermediate 9 6-Chloro-1-methyl-2H-3,1-benzoxazine-2,4(1H)-dione

As illustrated in the scheme above, NaH (2.43 g of a 60% dispersion, 60.8 mmol) was added to a solution of 6-chloro-2H-3,1-benzoxazine-2,4(1H)-dione (10.0 g, 50.6 mmol) dissolved in DMF (200 mL). The resulting mixture was stirred at room temperature for 30 min., and then methyl iodide (6.3 mL, 101 mmol) was added dropwise. After the reaction had stirred at room temperature overnight, it was concentrated in vacuo. Water and brine were added to the residue, and the aqueous layer was extracted with CH₂Cl₂ (2×). The combined organic phases were dried over Na₂SO₄, filtered, and concentrated in vacuo. The crude product was triturated with 3:1 hexanes:EtOAc. The solvent was removed by filtration, and the resulting solid was washed with additional 3:1 hexanes:EtOAc, followed by 100% hexanes. The product was dried briefly under vacuum to produce the title compound as a pale yellow solid. (8.59 g, 80%). ¹H-NMR (DMSO-d₆): δ 7.96 (d, J=2.6 Hz, 1H), 7.89 (dd, J=2.6 Hz, J=9.0 Hz, 1H), 7.48 (d, J=9.0 Hz, 1H), 3.45 (s, 3H). MS (ESI) (M+H)⁺=212.

Intermediate 10 7-Chloro-3,4-dihydro-1-methyl-1H-1,4-benzodiazepine-2,5-dione

As illustrated in the scheme above, a mixture of 6-chloro-1-methyl-2H-3,1-benzoxazine-2,4(1H)-dione (6.00 g, 28.4 mmol) and glycine (2.14 g, 28.4 mmol) in glacial acetic acid (72 mL) was heated at reflux for 4 h. The reaction was cooled and concentrated in vacuo. Water was added to the residue, and the mixture was cooled to 0° C. NaHCO₃ was added to adjust the pH of the aqueous layer to approximately 8, and then the aqueous layer was extracted with CH₂Cl₂ (3×). The combined organic phases were dried over Na₂SO₄, filtered, and concentrated in vacuo. The crude product was triturated with Et₂O, the solvent was removed by filtration, and the resulting solid was washed with additional Et₂O to provide the title compound as a slightly yellow solid (5.657 g, 89%). ¹H-NMR (CDCl₃): δ 7.88 (d, J=2.5 Hz, 1H), 7.57 (br s, 1H), 7.53 (dd, J=2.6 Hz, J=8.7 Hz, 1H), 7.19 (d, J=8.6 Hz, 1H), 3.84 (d, J=6.1 Hz, 2H), 3.39 (s, 3H). MS (ESI) (M+H)⁺=225.

Intermediate 11 5,7-Dichloro-1,3-dihydro-1-methyl-2H-1,4benzodiazepin-2-one

As illustrated in the scheme above, 7-chloro-3,4-dihydro-1-methyl-1H-1,4-benzodiazepine-2,5-dione (5.00 g, 22.3 mmol) was suspended in POCl₃ (100 mL) and heated at 100° C. for 30 min. The reaction was cooled and concentrated in vacuo. Traces of POCl₃ were removed by adding toluene and concentrating the mixture in vacuo (2×). The residue was dissolved in CH₂Cl₂, the solution was cooled to 0° C., and Et₃N (6.8 mL, 48.8 mmol) was added dropwise. The mixture was stirred for 1 h and allowed to slowly warm to room temperature, and was then concentrated in vacuo once again. The residue was purified by silica gel column chromatography (5:1 CH₂Cl₂:EtOAc+0.5% Et₃N) to provide the title compound as an orange solid (4.68 g, 86%). ¹H-NMR (CDCl₃): δ 7.79 (d, J=2.5 Hz, 1H), 7.55 (dd, J=2.4 Hz, J=8.9 Hz, 1H), 7.23 (d, J=8.8 Hz, 1H), 4.67 (br s, 1H), 3.72 (br s, 1H), 3.39 (s, 3H). MS (ESI) (M+H)⁺=243.

Intermediate 12 7-Chloro-5-(2,4-dimethoxy-5-pyrimidinyl)-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one

As illustrated in the scheme above and following General Procedure 5, (2,4-dimethoxy-5-pyrimidinyl)boronic acid (0.939 g, 5.10 mmol), Pd₂(dba)₃ (0.064 g, 0.07 mmol), dry KF (0.890 g, 15.3 mmol), 5,7-dichloro-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one (1.12 g, 4.61 mmol) and P(t-Bu)₃ (0.42 mL of a 10% solution in hexanes, 0.21 mmol) were combined and heated for 20 h. After workup, purification of the crude product by silica gel column chromatography (1:3 hexanes:EtOAc) provided the title compound as a pale orange solid (1.20 g, 75%). ¹H-NMR (CDCl₃): δ 8.50 (s, 1H), 7.49 (dd, J=2.4 Hz, J=8.8 Hz, 1H), 7.29 (d, J=8.8 Hz, 1H), 7.13 (d, J=2.4 Hz, 1H), 4.84 (d, J=10.8 Hz, 1H), 4.06 (s, 3H), 3.79 (s, 3H), 3.76 (d, J=10.8 Hz, 1H), 3.42 (s, 3H). MS (ESI) (M+H)⁺=347.

Intermediate 13 3-Azido-7-chloro-5-(2,4-dimethoxy-5-pyrimidinyl)-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one

As illustrated in the scheme above and following General Procedure 6, KHMDS (7.0 mL of 0.5 M in toluene, 3.5 mmol), 7-chloro-5-(2,4-dimethoxy-5-pyrimidinyl)-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one (1.16 g, 3.35 mmol), trisyl azide (2.60 g, 8.40 mmol) and acetic acid (0.85 mL, 14.8 mmol) were combined. After workup, purification of the crude product by silica gel column chromatography (2:3 hexanes:EtOAc) provided the title compound as a pale yellow solid (1.23 g, 95%). ¹H-NMR (CDCl₃): δ 8.64 (s, 1H), 7.54 (dd, J=2.3 Hz, J=8.8 Hz, 1H), 7.33 (d, J=8.8 Hz, 1H), 7.19 (d, J=2.3 Hz, 1H), 4.51 (s, 1H), 4.08 (s, 3H), 3.78 (s, 3H), 3.47 (s, 3H). MS (ESI) (M+H)⁺388.

Intermediate 14 3-Amino-7-chloro-5-(2,4-dimethoxy-5-pyrimidinyl)-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one

As illustrated in the scheme above and following General Procedure 7, 3-azido-7-chloro-5-(2,4-dimethoxy-5-pyrimidinyl)-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one (1.22 g, 3.15 mmol) and PS—PPh₃ (23.0 g of 1.37 mmol/g, 31.5 mmol) were combined. After workup and purification by “catch and release,” the title compound was obtained as a brown solid (1.15 g, quantitative). 1H-NM (CDCl₃): δ 8.56 (s, 1H), 7.50 (dd, J=2.4 Hz, J=8.9 Hz, 1H), 7.30 (d, J=8.8 Hz, 1H), 7.15 (d, J=2.5 Hz, 1H), 4.46 (s, 1H), 4.06 (s, 3H), 3.77 (s, 3H), 3.46 (s, 3H), 2.85-2.12 (br s, 2H). MS (ESI) (M+H)⁺=362.

Intermediate 15 7-Chloro-5-(2,4-dimethoxy-5-pyrimidinyl)-1,3-dihydro-3-isothiocyanato-1-methyl-2H-1,4-benzodiazepin-2-one

As illustrated in the scheme above and following General Procedure 2, 3-amino-7-chloro-5-(2,4-dimethoxy-5-pyrimidinyl)-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one (1.15 g, 3.18 mmol) and thiophosgene (0.49 mL, 6.4 mmol) were combined. Purification of the crude product by silica gel column chromatography (9:1 CH₂Cl₂:EtOAc) provided the title compound as a viscous dark yellow oil (0.560 g, 44%). ¹H-NMR (CDCl₃): δ 8.60 (s, 1H), 7.55 (dd, J=2.5 Hz, J=8.8 Hz, 1H), 7.33 (d, J=8.8 Hz, 1H), 7.17 (d, J=2.5 Hz, 1H), 5.16 (s, 1H), 4.07 (s, 3H), 3.78 (s, 3H), 3.49 (s, 3H). MS (ESI) (M+H)⁺=404.

Example 22 N-[7-chloro-5-(2,4-dimethoxy-5-pyrimidinyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzodiazepin-3-yl]-N′-[4-(diethylamino)-2-methylphenyl]-thiourea

As illustrated in the scheme above, a solution of 7-chloro-5-(2,4-dimethoxy-5-pyrimidinyl)-1,3-dihydro-3-isothiocyanato-1-methyl-2H-1,4-benzodiazepin-2-one (0.069 g, 0.146 mmol) and N⁴,N⁴-diethyl-2-methyl-1,4benzenediamine (0.029 g, 0.161 mmol) in (CH₂Cl)₂ (5.0 mL) was heated at 70° C. for 16 h. The reaction was cooled and concentrated in vacuo, and the residue was purified by silica gel column chromatography (7:3 CH₂Cl₂:EtOAc) to provide the title compound as a light orange solid (0.074 g, 87%). ¹H-NMR (CDCl₃): δ 8.64 (s, 1H), 7.52 (dd, J=2.5 Hz, J=8.8 Hz, 1H), 7.40 (s, 1H), 7.35 (d, J=7.8 Hz, 1H), 7.32 (d, J=9.0 Hz, 1H), 7.20 (d, J=2.2 Hz, 1H), 7.17-7.13 (m, 1H), 6.54-6.51 (m, 2H), 6.06 (d, J=8.0 Hz, 1H), 4.05 (s, 3H), 3.74 (s, 3H), 3.42 (s, 3H), 3.40-3.27 (m, 4H), 2.30 (s, 3H), 1.17 (t, J=7.1 Hz, 6H). HRMS calculated for (C₂₈H₃₂ClN₇O₃S+H) (M+H)⁺: 582.2054. Found (ESI): 582.2076.

Intermediate 16 7-Chloro-1,3-dihydro-1-methyl-5-(3-thienyl)-2H-1,4-benzodiazepin-2-one

As illustrated in the scheme above and following General Procedure 5,3-thienylboronic acid (1.15 g, 9.01 mmol), Pd₂(dba)₃ (0.113 g, 0.123 mmol), dry KF (1.57 g, 27.0 mmol), 5,7-dichloro-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one (1.98 g, 8.15 mmol) and P(t-Bu)₃ (0.75 mL of a 10% solution in hexanes, 0.37 mmol) were combined and heated for 16 h. After workup, purification of the crude product by silica gel column chromatography (9:1 CH₂Cl₂:EtOAc) provided the title compound as a yellow solid (1.44 g, 61%). ¹H-NMR (CDCl₃): δ 7.54-7.49 (m, 4H), 7.37 (dd, J=3.0 Hz, J=5.0 Hz, 1H), 7.29 (d, J=9.4 Hz, 1H), 4.76 (d, J=10.9 Hz, 1H), 3.77 (d, J=10.9 Hz, 1H), 3.38 (s, 3H). MS (ESI) (M+H)⁺=291.

Intermediate 17 3-Azido-7-chloro-1,3-dihydro-1-methyl-5-(3-thienyl)-2H-1,4-benzodiazepin-2-one

As illustrated in the scheme above and following General Procedure 6, KHMDS (10.4 mL of 0.5 M in toluene, 5.20 mmol), 7-chloro-1,3-dihydro-1-methyl-5-(3-thienyl)-2H-1,4-benzodiazepin-2-one (1.44 g, 4.95 mmol), trisyl azide (3.83 g, 12.4 mmol) and acetic acid (1.25 mL, 21.8 mmol) were combined. After workup, purification of the crude product by silica gel column chromatography (7:3 hexanes:EtOAc) provided the title compound as a pale yellow solid (1.60 g, 98%). ¹H-NMR (CDCl₃): δ 7.61-7.55 (m, 4H), 7.40 (dd, J=2.9 Hz, J=5.1 Hz, 1H), 7.33 (dd, J=1.0 Hz, J=8.2 Hz, 1H), 4.54 (s, 1H), 3.44 (s, 3H). MS (ESI) (M+H)⁺=332.

Intermediate 18 3-Amino-7-chloro-1,3-dihydro-1-methyl-5-(3-thienyl)-2H-1,4-benzodiazepin-2-one

As illustrated in the scheme above and following General Procedure 7, 3-azido-7-chloro-1,3-dihydro-1-methyl-5-(3-thienyl)-2H-1,4-benzodiazepin-2-one (1.60 g, 4.82 mmol) and PS—PPh₃ (30.0 g of 1.37 mmol/g, 41.1 mmol) were combined. After workup and purification by “catch and release,” the title compound was obtained as a brown solid (1.35 g, 92%). ¹H-NMR (CDCl₃): δ 7.56-7.50 (m, 4H), 7.36 (dd, J=3.1 Hz, J=4.9 Hz, 1H), 7.29 (d, J=8.8 Hz, 1H), 4.46 (s, 1H), 3.43 (s, 3H), 2.35-2.15 (br s, 2H). MS (ESI) (M+H)⁺=306.

Intermediate 19 7-Chloro-1,3-dihydro-3-isothiocyanato-1-methyl-5-(3-thienyl)-2H-1,4-benzodiazepin-2-one

As illustrated in the scheme above and following General Procedure 2,3-amino-7-chloro-1,3-dihydro-1-methyl-5-(3-thienyl)-2H-1,4-benzodiazepin-2-one (1.35 g, 4.41 mmol) and thiophosgene (0.67 mL, 8.8 mmol) were combined. Purification of the crude product by silica gel column chromatography (100% CH₂Cl₂) provided the title compound as a yellow solid (1.04 g, 68%). ¹H-NMR (CDCl₃): δ 7.61-7.54 (m, 4H), 7.39 (dd, J=2.9 Hz, J=5.3 Hz, 1H), 7.33 (d, J=8.8 Hz, 1H), 5.20 (s, 1H), 3.46 (s, 3H). MS (ESI) (M+H)⁺=348.

Example 23 N-[7-Chloro-2,3-dihydro-1-methyl-2-oxo-5-(3-thienyl)-1H-1,4-benzodiazepin-3-yl]-N′-[4-(4-morpholinyl)-1-naphthalenyl]thiourea

This reaction was carried out in a multiwell plate. As illustrated in the scheme above, aA mixture of 7-chloro-1,3-dihydro-3-isothiocyanato-1-methyl-5-(3-thienyl)-2H-1,4-benzodiazepin-2-one (156 μL of a 0.128 M solution in (CH₂Cl)₂, 0.020 mmol), 4-(4-morpholinyl)-1-naphthalenamine (44 μL of a 0.5 M solution in DMA, 0.022 mmol), and (CH₂Cl)₂ (300 μL) was agitated and heated at 70° C. for 22 h. The reaction was cooled and concentrated in vacuo, and the residue was redissolved in DMA (25 μL) and (CH₂Cl)₂ (275 μL). Polyamine resin HL (NovaBiochem) was added (20 mg of 4.53 mmol/g, 0.091 mmol), and the mixture was agitated at room temperature overnight. The resin was removed by filtration and washed with additional (CH₂Cl)₂ and MeOH. The filtrate was concentrated in vacuo to provide the title compound. MS (ESI) (M+H)⁺=576.

Intermediate 20 7-Chloro-1,3dihydro-1-methyl-5-(3-pyridinyl)-2H-1,4-benzodiazepin-2-one

As illustrated in the scheme above and following General Procedure 4,5,7-dichloro-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one (1.99 g, 8.19 mmol), Na₂CO₃ (0.868 g, 8.19 mmol), PdCl₂(dppf) (0.335 g, 0.410 mmol) and (3-pyridinyl)boronic acid (1.01 g, 8.19 mmol) were combined and heated for 14 h. After workup, purification of the crude product by silica gel column chromatography (100% EtOAc) provided the title compound (1.51 g, 64%). ¹H-NMR (CDCl₃): δ 8.78 (d, J=1.6 Hz, 1H), 8.72 (dd, J=1.6 Hz, J=4.8 Hz, 1H), 8.02 (dt, J=1.6 Hz J=8.0 Hz, 1H),7.56 (dd, J=2.4 Hz, J=8.8 Hz,1H), 7.39 (dd, J=4.8 Hz, J=8.0 Hz, 1H), 7.34 (d, J=8.8 Hz, 1H), 7.28 (d, J=2.4 Hz, 1H), 4.89 (d, J=10.4 Hz, 1H), 3.80 (d, J=10.8 Hz, 1H), 3.41 (s, 3H); MS (ESI) (M+H)⁺=286.

Intermediate 21 3-Azido-7-chloro-1,3-dihydro-1-methyl-5-(3-pyridinyl)-2H-1,4benzodiazepin-2-one

As illustrated in the scheme above and following General Procedure 6, KHMDS (10.5 mL of 0.5 M in toluene, 5.25 mmol), 7-chloro-1,3-dihydro-1-methyl-5-(3-pyridinyl)-2H-1,4-benzodiazepin-2-one (1.43 g, 4.99 mmol), trisyl azide (3.86 g, 12.5 mmol) and acetic acid (1.26 mL, 22.0 mmol) were combined. After workup, purification of the crude product by silica gel column chromatography (1:1 CH₂Cl₂:EtOAc) provided the title compound as a yellow foam (1.47 g, 90%). ¹H-NMR (CDCl₃): δ 8.80 (s, 1H), 8.76 (d, J=3.6 Hz, 1H), 8.14 (dt, J=1.6 Hz, J=8.0 Hz, 1H), 7.61 (dd, J=2.0 Hz, J=8.4 Hz, 1H), 7.44 (ddd, J=0.8 Hz, J=4.8 Hz, J=8.0 Hz, 1H), 7.38 (d, J=8.8 Hz, 1H), 7.35 (d, J=2.4 Hz, 1H), 4.56 (s, 1H), 3.48 (s, 3H). MS (ESI) (M+H)⁺=327.

Intermediate 22 3-Amino-7-chloro-1,3-dihydro-1-methyl-5-(3-pyridinyl)-2H-1,4-benzodiazepin-2-one

As illustrated in the scheme above and following General Procedure 7, 3-azido-7-chloro-1,3-dihydro-1-methyl-5-(3-pyridinyl)-2H-1,4-benzodiazepin-2-one (1.47 g, 4.49 mmol) and PS—PPh₃ (16.4 g of 1.37 mmol/g, 22.4 mmol) were combined. After workup and purification by “catch and release,” the title compound was obtained as a slightly brown solid (1.46 g, quantitative). ¹H-NMR (CDCl₃): δ 8.76 (d, J=2.4 Hz, 1H), 8.72 (dd, J=1.6 Hz, J=5.2 Hz, 1H), 8.06 (dt, J=2.4 Hz, J=7.6 Hz, 1H), 7.58 (dd, J=2.4 Hz, J=8.8 Hz, 1H), 7.40 (dd, J=4.8 Hz, J=8.0 Hz, 1H), 7.35 (d, J=8.8 Hz, 1H), 7.29 (d, J=2.4 Hz, 1H), 4.51 (s, 1H), 3.46 (s, 3H), 2.41 (br s, 2H). MS (ESI) (M+H)⁺=301.

Intermediate 23 7-Chloro-1,3-dihydro-3-isothiocyanato-1-methyl-5-(3-pyridinyl)-2H-1,4-benzodiazepin-2-one

As illustrated in the scheme above, a solution of 3-amino-7-chloro-1,3-dihydro-1-methyl-5-(3-pyridinyl)-2H-1,4benzodiazepin-2-one (1.36 g, 4.52 mmol) in dry THF (55 mL) was cooled to −15° C. Carbon disulfide (2.7 mL, 45 mmol) was added, followed by EDCI (1.73 g, 9.03 mmol). The mixture was stirred for 10 min., and then Et₃N (1.26 mL, 9.04 mmol) was added. The reaction was stirred for 16 h while it was allowed to slowly warm to room temperature. The precipitated solid was removed by filtration and was washed well with CH₂Cl₂ and then discarded. The filtrate was concentrated in vacuo, and the residue was dissolved in CH₂Cl₂. The organic phase was washed with water, saturated NaHCO₃, and brine, and was then dried over Na₂SO₄, filtered, and concentrated in vacuo. Purification of the crude product by silica gel column chromatography (3:1 CH₂Cl₂:EtOAc) provided the title compound as a solid (0.652 g, 42%). ¹H-NMR (CDCl₃): δ 8.75 (s, 2H), 8.12 (dt, J=2.0 Hz, J=8.0 Hz, 1H), 7.63 (dd, J=2.0 Hz, J=8.8 Hz, 1H), 7.43 (dd, J=4.8 Hz, J=8.0 Hz, 1H), 7.39 (d, J=8.8 Hz, 1H), 7.33 (d, J=2.4 Hz, 1H), 5.22 (s, 1H), 3.50 (s, 3H). MS (ESI) (M+H)⁺=343.

Example 24 N-[7-Chloro-2,3-dihydro-1-methyl-2-oxo-5-(3-pyridinyl)-1H-1,4-benzodiazepin-3-yl]-N′-[4-(4-morpholinyl)-1-naphthalenyl]thiourea

A solution of 7-chloro-1,3-dihydro-3-isothiocyanato-1-methyl-5-(3-pyridinyl)-2H-1,4-benzodiazepin-2-one (0.0282 g, 0.0823 mmol) and 4-(4-morpholinyl)-1-naphthalenamine (0.0188 g, 0.0823 mmol) in (CH₂Cl)₂ (2.5 mL) was heated at 70° C. for 24 h. The reaction was cooled and concentrated in vacuo, and the residue was purified by silica gel column chromatography (1:2 CH₂Cl₂:EtOAc) to provide the title compound (0.0263 g, 56%). ¹H-NMR (CDCl₃): δ 8.72-8.68 (m, 2H), 8.29-8.25 (m, 1H), 8.11-8.07 (m, 1H), 8.05-8.00 (m, 2H), 7.64-7.55 (m, 5H), 7.38-7.33 (m, 3H), 7.15 (d, J=8.0 Hz, 1H), 6.08 (d, J=7.6 Hz, 1H), 4.03-3.96 (br m, 4H), 3.39 (s, 3H), 3.15 (br s, 4H). HRMS calculated for (C₃₀H₂₇ClN₆O₂S+H) (M+H)⁺: 571.1683. Found (ESI): 571.1699. Anal. Calcd for C₃₀H₂₇ClN₆O₂S+0.7 H₂O: C, 61.73; H, 4.90; N, 14.40. Found: C, 61.93;H, 4.79; N, 13.87.

Intermediate 24 6-Fluoro-1-methyl-2H-3,1-benzoxazine-2,4(1H)-dione

As illustrated in the scheme above, NaH (0.312 g of a 60% dispersion, 7.80 mmol) was added to a solution of 6-fluoro-2H-3,1-benzoxazine-2,4(1H)-dione (1.176 g, 6.49 mmol) dissolved in DMF (60 mL). The resulting mixture was stirred at room temperature for 30 min., and then methyl iodide (0.81 mL, 13 mmol) was added dropwise. After the reaction had stirred at room temperature overnight, it was concentrated in vacuo. Water and brine were added to the residue, and the aqueous layer was extracted with CH₂Cl₂ (3×). The combined organic phases were dried over Na₂SO₄, filtered, and concentrated in vacuo. The crude product was triturated with Et₂O. The solvent was removed by filtration, and the resulting solid was washed with additional Et₂O. The product was dried briefly under vacuum to produce the title compound as a white solid (1.00 g, 79%). ¹H-NMR (CDCl₃): δ 7.86-7.82 (m, 1H), 7.55-7.48 (m, 1H), 7.22-7.17 (m, 1H), 3.61 (s, 3H). MS (ESI) (M+H)⁺=196.

Intermediate 25 7-Fluoro-3,4-dihydro-1-methyl-1H-1,4-benzodiazepine-2,5-dione

As illustrated in the scheme above, a mixture of 6-fluoro-1-methyl-2H-3,1-benzoxazine-2,4(1H)-dione (1.00 g, 5.12 mmol) and glycine (0.385 g, 5.13 mmol) in glacial acetic acid (13 mL) was heated at reflux for 4 h. The reaction was cooled and concentrated in vacuo. Water was added to the residue, and the mixture was cooled to 0° C. NaHCO₃ was added to adjust the pH of the aqueous layer to approximately 8, and then the aqueous layer was extracted with CH₂Cl₂ (4×). The combined organic phases were dried over Na₂SO₄, filtered, and concentrated in vacuo. The crude product was triturated with Et₂O, the solvent was removed by filtration, and the resulting solid was washed with additional Et₂O to provide the title compound as a slightly yellow solid (0.560 g, 52%). ¹H-NMR (DMSO-d₆): δ 8.80 (br s, 1H), 7.49-7.38 (m, 3H), 3.76 (br d, 1H), 3.47 (br d, 1H), 3.26 (s, 3H). MS (ESI) (M+H)⁺=209.

Intermediate 26 5-Chloro-7-fluoro-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one

As illustrated in the scheme above, 7-fluoro-3,4-dihydro-1-methyl-1H-1,4-benzodiazepine-2,5-dione (0.495 g, 2.38 mmol) was suspended in POCl₃ (10.6 mL) and heated at 100° C. for 30 min. The reaction was cooled and concentrated in vacuo. Traces of POCl₃ were removed by adding toluene and concentrating the mixture in vacuo (2×). The residue was dissolved in CH₂Cl₂, the solution was cooled to 0° C., and Et₃N (0.75 mL, 5.4 mmol) was added dropwise. The mixture was stirred for 0.5 h and allowed to slowly warm to room temperature, and was then concentrated in vacuo once again. The residue was purified by silica gel column chromatography (9:1 CH₂Cl₂:EtOAc+0.5% Et₃N) to provide the title compound as a light tan solid (0.422 g, 78%). ¹H-NMR (CDCl₃): δ 7.53-7.48 (m, 1H), 7.34-7.24 (m, 2H), 4.66 (br s, 1H), 3.72 (br s, 1H), 3.39 (s, 3H). MS (ESI) (M+H)⁺=227.

Intermediate 27 7-Fluoro-1,3-dihydro-1-methyl-5-phenyl-2H-1,4-benzodiazepin-2-one

As illustrated in the scheme above and following General Procedure 4, 5-chloro-7-fluoro-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one (0.201 g, 0.887 mmol), Na₂CO₃ (0.0940 g, 0.887 mmol), PdCl₂(dppf) (0.0362. g, 0.0443 mmol) and phenylboronic acid (0.108 g, 0.886 mmol) were combined and heated for 13 h. After workup, purification of the crude product by silica gel column chromatography (3:1 CH₂Cl₂:EtOAc) provided the title compound (0.180 g, 76%). ¹H-NMR (CDCl₃): δ 7.64-7.60 (m, 2H), 7.51-7.45 (m, 1H), 7.44-7.39 (m, 2H), 7.34 (dd, J=4.8 Hz, J=9.2 Hz, 1H), 7.31-7.25 (m, 1H), 7.02 (dd, J=2.8 Hz, J=8.8 Hz, 1H), 4.84 (d, J=10.8 Hz, 1H), 3.78 (d, J=10.8 Hz, 1H), 3.40 (s, 3H). MS (ESI) (M+H)⁺=269.

Intermediate 28 3-Azido-7-fluoro-1,3-dihydro-1-methyl-5-phenyl-2H-1,4-benzodiazepin-2-one

As illustrated in the scheme above and following General Procedure 6, KHMDS (1.34 mL of 0.5 M in toluene, 0.670 mmol), 7-fluoro-1,3-dihydro-1-methyl-5-phenyl-2H-1,4-benzodiazepin-2-one (0.171 g, 0.637 mmol), trisyl azide (0.493 g, 1.59 mmol) and acetic acid (0.16 mL, 2.8 mmol) were combined. After workup, purification of the crude product by silica gel column chromatography (100% CH₂Cl₂ to 9:1 CH₂Cl₂:EtOAc) provided the title compound as a pale yellow solid (0.163 g, 83%). ¹H-NMR (CDCl₃): δ 7.72-7.67 (m, 2H), 7.54-7.49 (m, 1H), 7.47-7.42 (m, 2H), 7.39 (dd, J=4.8 Hz, J=9.2 Hz, 1H), 7.36-7.30 (m, 1H), 7.08 (dd, J=2.8 Hz, J=8.4 Hz, 1H), 4.55 (s, 1H), 3.45 (s, 3H). MS (ESI) (M+H)⁺=310.

Intermediate 29 3-Amino-7-fluoro-1,3-dihydro-1-methyl-5-phenyl-2H-1,4-benzodiazepin-2-one

As illustrated in the scheme above and following General Procedure 7, 3-azido-7-fluoro-1,3-dihydro-1-methyl-5-phenyl-2H-1,4-benzodiazepin-2-one (0.0826 g, 0.267 mmol) and PS—PPh₃ (1.63 g of 1.64 mmol/g, 2.67 mmol) were combined. After workup and purification by “catch and release,” the title compound was obtained as a slightly yellow solid (0.0450 g, 59%). ¹H-NMR (CDCl₃): δ 7.65-7.60 (m, 2H), 7.51-7.46 (m, 1H), 7.44-7.39 (m, 2H), 7.36 (dd, J=4.8 Hz, J=9.2 Hz, 1H), 7.33-7.26 (m, 1H), 7.03 (dd, J=2.4 Hz, J=8.4 Hz, 1H), 4.48 (s, 1H), 3.45 (s, 3H), 2.30-1.60 (br s, 2H). MS (ESI) (M+H)⁺=284.

Example 25 N-(7-Fluoro-2,3-dihydro-1-methyl-2-oxo-5-phenyl-1H-1,4-benzodiazepin-3-yl)-N′-[2-methyl-4-(4-morpholinyl)phenyl]thiourea

As illustrated in the scheme above, a solution of 3-amino-7-fluoro-1,3-dihydro-1-methyl-5-phenyl-2H-1,4-benzodiazepin-2-one (0.0056 g, 0.020 mmol) and 4-(4-isothiocyanato-3-methylphenyl)morpholine (0.0048 g, 0.020 mmol) in DMA (0.5 mL) was heated at 70° C. for 16 h. The reaction was cooled and concentrated in vacuo, and the residue was lyophilized to provide the title compound (0.0104 g, quantitative). ¹H-NMR (CDCl₃): δ 7.63-7.58 (m, 2H), 7.50-7.45 (m, 1H), 7.44-7.36 (m, 4H), 7.36-7.26 (m, 3H), 7.09 (dd, J=2.4 Hz, J=8.4 Hz, 1H), 6.85-6.78 (m, 2H), 6.09-6.05 (m, 1H), 3.90-3.83 (br m, 4H), 3.41 (s, 3H), 3.21-3.15 (br s, 4H), 2.36 (s, 3H). MS (ESI) (M+H)⁺=518.HRMS calculated for (C₂₈H₂₈FN₅O₂S+H) (M+H)⁺: 518.2026. Found (ESI): 518.2117.

Intermediate 30 7-Chloro-1,3-dihydro-5-(6-methoxy-2-pyridinyl)-1-methyl-2H-1,4-benzodiazepin-2-one

As illustrated in the scheme above, absolution of 2-bromo-6-methoxypyridine (0.022 mL, 0.18 mmol) in dry Et₂O (0.3 mL) was added to a solution of n-BuLi (0.12 mL of 1.6 M in hexanes, 0.19 mmol) maintained at −40° C. The reaction was stirred at −40° C. for 20 min., and then B(OMe)₃ (0.022 mL, 0.19 mmol) was added dropwise. The reaction was stirred at −40° C. for 30 min., and then at room temperature for 3.5 h. The reaction was concentrated in vacuo, anhydrous MeOH was added to the residue, and the reaction was concentrated in vacuo once again. Dry DME (0.8 mL), 5,7-dichloro-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one (0.0392 g, 0.161 mmol), Pd(PPh₃)₄ (0.0093 g, 0.0080 mmol) and CsF (0.0612 g, 0.403 mmol) were added to the residue, and the mixture was heated to reflux for 15 h. Water (5 mL) and CH₂Cl₂ (5 mL) were added to the reaction mixture, and the layers were separated. The aqueous phase was extracted with additional CH₂Cl₂ (3×), and the combined organic phases were dried over Na₂SO₄, filtered, and concentrated in vacuo. Purification of the crude product by silica gel column chromatography (2:1 CH₂Cl₂:EtOAc) provided the title compound (0.029 g, 57%). ¹H-NMR (CDCl₃): δ 7.77 (d, J=7.6 Hz, 1H), 7.71-7.64 (m, 2H), 7.49 (dd, J=2.4 Hz, J=8.8 Hz, 1H), 7.27 (d, J=8.8 Hz, 1H), 6.83 (d, J=8.0 Hz, 1H), 4.85 (d, J=10.8 Hz, 1H), 3.86 (d, J=10.4 Hz, 1H), 3.81 (s, 3H), 3.39 (s, 3H). MS (ESI) (M+H)⁺=316.

Intermediate 31 3-Azido-7-chloro-1,3-dihydro-5-(6-methoxy-2-pyridinyl)-1-methyl-2H-1,4-benzodiazepin-2-one

As illustrated in the scheme above and following General Procedure 6, KHMDS (0.19 mL of 0.5 M in toluene, 0.095 mmol), 7-chloro-1,3-dihydro-5-(6-methoxy-2-pyridinyl)-1-methyl-2H-1,4-benzodiazepin-2-one (0.0290 g, 0.0918 mmol), trisyl azide (0.0710 g, 0.229 mmol) and acetic acid (0.023 mL, 0.40 mmol) were combined. After workup, purification of the crude product by silica gel column chromatography (49:1 CH₂Cl₂:EtOAc) provided the title compound (0.0204 g, 62%). ¹H-NMR (CDCl₃): δ 7.95 (d, J=7.2 Hz, 1H), 7.78-7.70 (m, 2H), 7.54 (dd, J=2.4 Hz, J=8.8 Hz, 1H), 7.31 (d, J=8.8 Hz, 1H), 6.87 (d, J=8.4 Hz, 1H), 4.64 (s, 1H), 3.80 (s, 3H), 3.44 (s, 3H). MS (ESI) (M+H)⁺=357.

Intermediate 32 3-Amino-7-chloro-1,3-dihydro-5-(6-methoxy-2-pyridinyl)-1-methyl-2H-1,4-benzodiazepin-2-one

As illustrated in the scheme above, a suspension of Pd/C (0.005 g of 10% on C) in MeOH (0.5 mL) under N₂ was treated with ammonium formate (0.0252 g, 0.400 mmol). The mixture was stirred for 10 min. and then transferred via Pasteur pipette to a suspension of 3-azido-7-chloro-1,3-dihydro-5-(6-methoxy-2-pyridinyl)-1-methyl-2H-1,4-benzodiazepin-2-one (0.0204 g, 0.0572 mmol) in MeOH (1.2 mL). The resulting mixture was stirred at room temperature for 3 h and was then filtered through a small pad of Celite. The filtrate was concentrated in vacuo, and the residue was purified by silica gel column chromatography (100% EtOAc, followed by 4:1 CH₂Cl₂:EtOAc) to provide the title compound (0.0148 g, 78%). ¹H-NMR (CDCl₃): δ 7.82 (dd, J=0.8 Hz, J=7.6 Hz, 1H), 7.71-7.66 (m, 2H), 7.51 (dd, 2.4 Hz, J=8.8 Hz, 1H), 7.28 (d, J=8.8 Hz, 1H), 6.83 (dd, J=0.8 Hz, J=8.4 Hz, 1H), 4.56 (s, 1H), 3.80 (s, 3H), 3.44 (s, 3H), 2.53 (br s, 2H). MS (ESI) (M+H)⁺=331.

Example 26 N-[7-Chloro-2,3-dihydro-5-(6-methoxy-2-pyridinyl)-1-methyl-2-oxo-1H-1,4-benzodiazepin-3-yl]-N′-[2-methyl-4-(4-morpholinyl)phenyl]thiourea

As illustrated in the scheme above, a solution of 3-amino-7-chloro-1,3-dihydro-5-(6-methoxy-2-pyridinyl)-1-methyl-2H-1,4-benzodiazepin-2-one (0.0113 g, 0.0342 mmol) and 4-(4-isothiocyanato-3-methylphenyl)morpholine (0.0080 g, 0.034 mmol) in (CH₂Cl)₂ (0.9 mL) was heated at 70° C. for 20 h. The reaction was cooled and concentrated in vacuo, and the residue was purified by silica gel column chromatography (2:1 CH₂Cl₂:EtOAc) to provide the title compound (0.0172 g, 89%) as a slightly yellow solid. ¹H-NMR (CDCl₃): δ 7.79 (dd, J=0.8 Hz, J=7.2 Hz, 1H), 7.70-7.65 (m, 2H), 7.54 (dd, J=2.4 Hz, J=8.8 Hz, 1H), 7.42 (br s, 1H), 7.30 (d, J=8.8 Hz, 1H), 7.28-7.25 (m partially hidden under CHCl₃, 2H), 6.85-6.80 (m, 3H), 6.14 (m, 1H), 3.88-3.85 (m, 4H), 3.78 (s, 3H), 3.40 (s, 3H), 3.22-3.18 (m, 4H), 2.36 (s, 3H). MS (M+H⁺=565. 

1. A compound of Formula (I), or pharmaceutically acceptable salt, diastereomer, or enantiomer thereof:

wherein R¹ is selected from optionally substituted acyl, optionally substituted alkyloxycarbonyl, optionally substituted alkyl, optionally substituted heteroalkyl, optionally substituted cycloalkyl, optionally substituted naphthyl, optionally substituted isoquinolyl, optionally substituted acridinyl, optionally substituted coumarinyl, optionally substituted carbazolyl, optionally substituted heterocyclyl, optionally substituted aryl-C₁₋₆alkyl, and optionally substituted heterocyclyl-C₁₋₆alkyl; X is —NHC(═S)N(R²)—; R³ is substituted aryl, optionally substituted C₁₋₁₂alkyl, optionally substituted C₃₋₁₂cycloalkyl, or optionally substituted heterocyclyl; R⁴ is, at each position, independently halogen, optionally substituted alkyl, optionally substituted heteroalkyl, nitro, cyano, hydroxy, —OR⁶, —SR⁶, —S(═O)R⁶, —S(═O)₂R⁶, —C(═O)R⁶, —C(═S)R⁶, —NR⁷R⁶, —C(═O)NR⁷R⁶, —NR⁷C(═O)R⁶, —SO₂NR⁷R⁶, —NR⁷SO₂R⁶, or —C(═O)OR⁶; R⁵, R⁶ and R⁷ are independently —H, or optionally substituted C₁₋₆alkyl; and R² is selected from —H, optionally substituted C₁₋₁₂alkyl, optionally substituted C₁₋₁₂heteroalkyl, substituted aryl, and optionally substituted heterocyclyl.
 2. The compound of claim 1, or pharmaceutically acceptable salt, diastereomer, or enantiomer thereof wherein: R¹ is optionally substituted naphthyl, optionally substituted isoquinolyl, optionally substituted acridinyl, optionally substituted coumarinyl, or optionally substituted carbazolyl, wherein said naphthyl, isoquinolyl, acridinyl, coumarinyl, and carbazolyl are optionally substituted by C₁₋₆alkyl , C₁₋₆heterocyclyl or amino; R² is —H or C₁₋₃alkyl; R³ is substituted aryl, optionally substituted heteroaryl or optionally substituted cycloalkyl; R⁴ is halogen, or C₁₋₃alkyl; and R⁵ is C₁₋₃alkyl.
 3. The compound of claim 2, or pharmaceutically acceptable salt, diastereomer, or enantiomer thereof, wherein: R¹ is optionally substituted naphthyl or optionally substituted isoquinolyl, wherein said naphthyl, and isoquinolyl are optionally substituted by C₁₋₆alkyl, C₁₋₆heterocyclyl or amino; R² is —H or C₁₋₃alkyl; R³ is optionally substituted cyclohexyl, optionally substituted pyridyl, optionally substituted thienyl, or optionally substituted pyrimidinyl, wherein said cyclohexyl, pyridyl, thienyl, and pyrimidinyl are optionally substituted by halogen, methoxy, or C₁₋₃alkyl; R⁴ is halogen; and R⁵is methyl.
 4. The compound of claim 1, or pharmaceutically acceptable salt, diastereomer, or enantiomer thereof, wherein: R¹ is optionally substituted naphthyl or, optionally substituted isoquinolyl, wherein said naphthyl, and isoquinolyl are optionally substituted by C₁₋₆alkyl, C₁₋₆heterocyclyl or amino; R² is C₁₋₃alkyl; R³ is optionally substituted cyclohexyl, substituted phenyl, substituted pyridyl, substituted thienyl, or substituted pyrimidinyl, wherein said cyclohexyl is optionally substituted by halogen, methoxy, or C₁₋₃alkyl; R⁴ is halogen; and R⁵is methyl.
 5. The compound of claim 1, or pharmaceutically acceptable salt, diastereomer, or enantiomer thereof, wherein R⁵ is —H or substituted C₁₋₆alkyl.
 6. The compound of claim 1, or pharmaceutically acceptable salt, diastereomer, or enantiomer thereof, wherein: R¹ is optionally substituted naphthy or optionally substituted isoquinolyl, wherein said naphthyl, and isoquinolyl are optionally substituted by C₁₋₆alkyl, C₁₋₆heterocyclyl or amino; R² is —H or C₁₋₃alkyl; R³ is optionally substituted cyclohexyl, substituted phenyl, substituted pyridyl, substituted thienyl, or substituted pyrimidinyl, wherein said cyclohexyl is optionally substituted by halogen, methoxy, or C₁₋₃alkyl; R⁴ is halogen; and R⁵ is substituted C₁₋₃alkyl.
 7. A compound of Formula I, or pharmaceutically acceptable salt, diastereomer, or enantiomer thereof:

wherein: R¹ is optionally substituted phenyl, optionally substituted naphthyl, optionally substituted isoquinolyl, optionally substituted acridinyl, optionally substituted coumarinyl, optionally substituted carbazolyl, or a first divalent group selected from optionally substituted C₁₋₁₂alkylene and optionally substituted C₁₋₁₂heteroalkylene; wherein said naphthyl, isoquinolyl, acridinyl, coumarinyl, and carbazolyl are optionally substituted by C₁₋₆alkyl, C₁₋₆heterocyclyl or amino, wherein said C₁₋₁₂alkylene and C₁₋₁₂heteroalkylene are optionally substituted by C₁₋₆alkyl, aryl-C₁₋₆alkyl, aryl or heterocyclyl; X is —NHC(═S)N(R²)—; R² is —H, C₁₋₃alkyl, or a second divalent group selected from a single bond, an optionally substituted alkylene and an optionally substituted heteroalkylene; wherein said second divalent group together with said first divalent group forms a portion of a ring; R³ is substituted aryl, optionally substituted heteroaryl or optionally substituted cycloalkyl; R⁴ is halogen, or C₁₋₃alkyl; and R⁵ is C₁₋₃alkyl.
 8. The compound of claim 7, or pharmaceutically acceptable salt, diastereomer, or enantiomer thereof, wherein: R¹ is optionally substituted naphthyl, optionally substituted isoquinolyl, wherein said naphthyl, and isoquinolyl are optionally substituted by C₁₋₆alkyl, C₁₋₆heterocyclyl or amino; R¹ is —H or C₁₋₃alkyl; R³ is optionally substituted cyclohexyl, substituted phenyl, substituted pyridyl, substituted thienyl, or substituted pyrimidinyl, wherein said cyclohexyl is optionally substituted by halogen, methoxy, or C₁₋₃alkyl; R⁴ is halogen; and R⁵ methyl.
 9. The compound of claim 1, or pharmaceutically acceptable salt, diastereomer, or enantiomer thereof, wherein: R¹ is optionally substituted naphthyl, optionally substituted isoquinolyl, wherein said naphthyl and isoquinolyl are optionally substituted by C₁₋₆alkyl, C₁₋₆heterocyclyl or amino; R² is —H or C₁₋₃alkyl; R³ is optionally substituted cyclohexyl, substituted phenyl, substituted pyridyl, substituted thienyl, or substituted pyrimidinyl, wherein said cyclohexyl is optionally substituted by halogen, methoxy, or C₁₋₃alkyl; R⁴ is halogen; and R⁵ methyl.
 10. A mixture of two or more compounds selected from a compound of claim 1, a pharmaceutically acceptable salt thereof, a diasteromer thereof, and an enantiomer thereof.
 11. A compound of Formula I, or pharmaceutically acceptable salt, diastereomer, or enantiomer thereof:

wherein; —X—R¹, in combination, is selected from —NHC(═S)N(R²)(R¹) and

R¹ is optionally substituted phenyl, optionally substituted naphthyl, optionally substituted isoquinolyl, wherein said phenyl, naphthyl and isoquinolyl are optionally substituted by C₁₋₆alkyl, C₁₋₆heterocyclyl or amino; R² is —H, or C₁₋₃alkyl;

is a nitrogen containing heterocyclyl, which is optionally substituted by one or more —R⁸, and which includes a bond on the nitrogen that links to other group of formula (I); R⁸ is —H, optionally substituted aryl, optionally substituted heterocyclyl, optionally substituted C₁₋₆alkyl, —OH, or C₁₋₆alkoxy, wherein R⁸ is optionally fused with the ring of

R³ is optionally substituted C₃₋₁₂cyclohexyl, substituted phenyl, optionally substituted pyridyl, optionally substituted thienyl, or optionally substituted pyrimidinyl, wherein said C₃₋₁₂cyclohexyl, phenyl, pyridyl, thienyl and pyrimidinyl are optionally substituted by halogen, methoxy, or C₁₋₃alkyl; R⁴ is halogen; and R⁵ is methyl.
 12. A compound as claimed in claim 11, wherein said nitrogen containing heterocyclyl is selected from piperazinyl, morpholinyl, piperidyl, and pyrrolidinyl. 